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Received: 15 January 2019    Revised: 18 February 2019    Accepted: 19 February 2019

DOI: 10.1111/ijlh.13006

SUPPLEMENT ARTICLE

The wonderful story of monoclonal antibodies

Marie C Béné

Nantes University Hospital, CRCINA,

Nantes, France Abstract
Monoclonal antibodies have become daily partners of both biologists and clinicians,
Correspondence
Hematology Biology, Nantes University as reagents and therapeutic agents. Behind their odd names and incredible diversity
Hospital, Nantes, France. lies an amazing story of inventiveness and daring. This review tries to retrace the
Email: [email protected]
major steps of this saga, initiated by the search for anti-­rabbit red blood cell anti­
bodies and currently culminating in amazing molecular constructions saving lives.
After some historical and basic reminders, the fields of reagents and drugs will be
addressed. This invaluable contribution of immunology to the understanding of both
physiology and treatment clearly deserves to be fully recognized.

KEYWORDS
biologics, flow cytometry, immunoglobulins, monoclonal antibodies, pathology

1 | A NTI B O D I E S O R I M M U N O G LO B U LI N S ? experiment reported in 1891 by Guido Tizzoni and Giuseppina

Cattani.5 They concluded from their experiments that these sub-
The discovery of soluble proteins in the plasma/serum able to react stances were “globulins.” Almost 50 years later, after the develop-
against foreign material only occurred at the end of the XIXth cen- ment of electrophoretic techniques, Arne Tiselius identified the
tury. Emil Adolf von Behring and Shibasaburo Kitasato, in 1890, presence of antibodies in one of the fractions separated in serum
demonstrated how small doses of tetanus or diphtheria “toxoids” by this methodology.6 These fractions were attributed Greek letters
administered to animals led to the formation of a protective sub- and antibodies then became synonyms to “gammaglobulins,” further
stance in the ­animals’ serum that could be effectively transferred studied notably by Gerald Maurice Edelman,7 a feat for which he
to recipients.1 Because of the function or type of reaction that oc- received the Nobel Prize together with Rodney Porter in 1972. The
curred between these elusive molecules and various types of exog- rest of the story is quite fuzzy, until a nomenclature including the
enous substances, they were first called “antitoxins,” “precipitins,” word “immunoglobulins” was finally agreed upon,8 that word having
“agglutinins” or even “reagins.” Paul Ehrlich seems to have been the been coined by Joseph F Heremans in 1960.9
first to coin the German word “Antikörper” in 1890, but it took some The structure of antibodies also has an interesting history, ini-
2
time for it to be accepted and commonly used worldwide. Basically, tiated by the “side chain” theory of Paul Ehrlich in 1901.10 This
this term is quite confusing, by describing either a “body that rec- totally empirical concept was incredibly accurate, imagining that
ognizes something” or “something that recognizes a body”. The latter some cells carried the so-­
c alled side chains, specific for given
acceptation was finally adopted after clarification by Maurice Nicolle structures, at that time diphtheria or tetanus toxins. He proposed
in 1907,3 together with the notion that this “body” was an antigen. that these side chains could be released in the serum when the
The word “antigen” itself has a complicated story and was seemingly structures they recognized entered the animal. Before this, having
initially proposed by Lazslo Deutsch when he was working in Paris become head of the Institute of Serum Research and Testing in
at the Pasteur Institute in the laboratory of lja Iljitsch Metschnikow, Berlin, he applied the immunization protocols he had developed
in 1899. 4 with Koch and von Behring.11 He was the one who proposed the
Antitoxins were the first to be identified as molecules that could use of horses for the standardized commercial production of im-
precipitate when serum was mixed with magnesium sulfate, in an mune serum. In the golden age of serotherapy, countless brave

8  |  wileyonlinelibrary.com/journal/ijlh
© 2019 John Wiley & Sons Ltd Int J Lab Hematol. 2019;41(Suppl. 1):8–14.
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TA B L E   1   Proposed terminology for immunoglobulin fragments, adapted from the nomenclature of human immunoglobulins, Immunology
1963

Proposed terminology Fab fragment Fc fragment Fd fragment F(ab’)² Fab’

Former terminology I;II III A piece 5S peptic Univalent peptide

fragment fragment

horses and rabbits have to be commended for having produced Biochemical studies, using mostly crystallography and sequenc-
protective sera, rich in specific immunoglobulins, that, among ing, meanwhile tried to better understand the structure of these gly-
other feats, saved soldiers of World wars from tetanus, diphtheria coproteins. The striking juxtaposition of molecular motifs of about
or other ailments. 100 amino acids including cysteines strategically placed to allow
The era of serotherapy blossomed and progressed. Many re- the formation of disulfide bonds was progressively identified as an
12
searchers, Edwin Cohn among others, devised ways to purify extremely clever structure. Further modelization highlighted the
human immunoglobulins in order to provide patients that needed systematic organization of these amino acids in antiparallel beta-­
it with passive immunization. This led to precious preparations di- pleated sheets linked by loose loops. The notion of domain emerged.
rected toward infectious agents, helpful to alleviate infections in The fact that this molecular structure allowed for adhesion between
immunocompromised/immunodeficient patients. Broader prepara- siblings and was in fact used by quite a number of transmembrane
tions of IVIG or intravenous immunoglobulins were developed, still or extracellular glycoproteins progressively led to the notion of “im-
widely used not only for immunodeficient patients but also for the munoglobulin superfamily”. 20 Indeed, the unique assembly of these
treatment of such autoimmune disorders as immune thrombocyto- domains confers them with a striking capacity to interact together.
13
penic purpura. Nowadays, IVIG are processed in order to try and However, even though these notions became increasingly clear,
limit the adverse reactions they may yield.14 the exquisite specificity of immunoglobulins for a given antigen, al-
though clearly carried by their variable domains, remained a mystery.

2 | D I S COV E RY O F A N A M A Z I N G
M O LEC U L A R S TRU C T U R E 3 | R I S E O F M O N O C LO N A L A NTI B O D I E S

But much work and time were still needed to unravel the mysteries In the 1970s, this question of variability was gnawing at the brilliant
of immunoglobulins. This was helped by refining animal immuniza- mind of Cesar Milstein, a researcher of Argentinian origin, work-
tion techniques but also by the characterization of myelomatous ing in London in the renowned laboratory of Frederick Sanger. The
proteins from patients with multiple myeloma. By the time the WHO model developed by Cesar Milstein was that of rabbits immunized
nomenclature was published in the early 60s,8 the basic structure with sheep red blood cells. His tools were antibody purification,
of immunoglobulins had been deciphered, a model of the associa- crystallography, then RNA sequencing, but he was puzzled by the
tion of heavy and light chains having been proposed in 1962 by differences observed each time he was using a new batch of rab-
Rodney Porter.15 Heavy chain isotypes had been described for IgG bit antiserum. Because he believed that somatic mutations were
and IgM in the 1940s, IgA in 1956, and IgD in 1964. IgE would have responsible for the generation of specific immunoglobulins, he em-
to wait until 1967 to be characterized and linked to the long-­known barked on long-­term culture of cell lines or myeloma cells. He thus
“reagins.” The light chains of immunoglobulins had been initially de- participated to experiments conducted on hybridomas generated
scribed as proteins present in the urine of myelomatous patients by with Abraham Karpas and Dick Cotton, following observations by
Henry Bence Jones as early as 1864.16 That there were two types of Hervé Bazin in Belgium. Immortalization of a rat myeloma with a
Bence Jones proteins was however only elucidated almost a century mouse myeloma after fusion by the Sendai virus resulted in a hybri-
later, in 1950 by Korngold and Lipari who used the first letter of their doma producing both types of myelomatous immunoglobulins. After
respective names to give them their Greek equivalent denomination identification of the two respective RNAs in the cells, the conclusion
of kappa and lambda chains.17 Further chemical and enzymatic ex- was that each had brought, from its nucleus, the genetic message of
periments allowed to discover that the four-­chain structure could each immunoglobulin. The big problem was that these immunoglob-
be subdivided in fragments of different functions (Table 1), their ulins had no known reactivity toward a recognized antigen. After a
nomenclature being also clarified in 19648 mostly as Fc (for crystal- talk he gave in the Basel Institute, he was approached by a young fel-
lizable) and Fab (for antigen-binding sometimes also referred to as low named George Kohler, fascinated by the topic and who applied
antibody fragment). to come as a postdoctoral fellow in Cambridge. There, as an ancil-
In the 1960s, several seminal works also first identified that lym- lary project, Milstein asked him if he could devise a way to obtain a
phocytes were the cells responsible for specific immunity,18 then steadily similar batch of anti-­sheep red blood cell immunoglobulins.
that the production of antibodies was the realm of B cells.19 Aware of the hybridoma experiment, Kohler grew the idea to fusion
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not two myelomas but B cells from an immunized animal (with sheep were published in a mythic book in 1984. 22 Thanks to the statisti-
red blood cells) and an immortal myeloma. The screen for hybrido- cal gifts of Catherine Hill at Gustave Roussy cancer center, results
mas producing one single species of anti-­sheep red blood cells im- from 55 research groups were computed to group within clusters the
munoglobulins was successful. The cells were growing as a single 139 antibodies provided by different laboratories. All antibodies in a
clone, indefinitely, and secreted unlimited amounts of the immuno- cluster displayed the same reactivity. This statistical approach is the
globulin encoded by their DNA: monoclonal antibodies! Milstein was one that yielded the name “Cluster of differentiation” or CD. Because
first thrilled to have a tool allowing to test (unsuccessfully) his theory so many of the new “antigens” discovered were utterly unknown,
of somatic mutations. However, the two scientists soon recognized they were attributed the same name. CD is thus a denomination
that they had something much bigger, allowing to produce “à la carte” characterizing monoclonal antibodies of the same specificity or the
antibodies. molecule thus identified. The first HLDA confirmed 11 specificities
and proposed four additional provisional ones, numbered from CD1
to CDw15. After the 10th, in 2014 in Barcelona, 371 CD have been
4 |  M O N O C LO N A L A NTI B O D I E S A S N E W formally recognized, quite a number of them being subdivided in
R E AG E NT S subcluster isoforms. 26 Upon regular international meetings, the rules
were refined, requiring at least two antibodies from two different
Kohler and Milstein, after a rebuttal in 1975 from the journal Science, labs to have been reported, together with the protein and later gene
finally published their outbreaking results in a short letter in Nature structure of the related molecule. The production of these reagents
21
in August of the same year. This initiated a revolution and much left mice peritoneum to reach the more ethical fermenters allow-
ebullition in the immunological world. The technique by itself was ing for mass production. Along the way, however, some extremely
rather simple. It involved immunization of mice with small doses of precious hybridomas were lost and some were never recognized for
antigen, removal of their spleen, and mixture of splenic B-cells with their possible value. Indeed, we still are far away from of the ex-
a rat myeloma cell line. Sendai virus, then polyethylene glycol, were pected 7000 cell proteins that could be identified by such tools. 27 Of
found to achieve the random hybridization of cells. Use of a selective note, some monoclonal antibodies have also largely been produced
medium allowed only hybridomas of a B-­lymphocyte and a myeloma toward already known structures of which they did not change the
cell to survive. The trickiest part, probably, was the tedious seeding name. They, however, provided exquisite tools to study them.
of limited dilutions of the hybridomas, drop by drop in multiple-well Monoclonal antibodies were first used as ascites fluids as men-
plates, with the aim of having no more than one cell in each well. tioned above. The monospecificity and huge affinity of the clonal
After selecting wells seeded with hybridomas and letting the lat- immunoglobulins allowed for extensive research, based on the high
ter grow, another tedious task was to seek for the supernatant(s) level of dilution permitted for experiments in many types of settings,
containing immunoglobulins of the expected specificity. Afterwards, from immunochemistry and immunophenotyping to immunohistol-
mice came back in the picture in the now abandoned peritoneal in- ogy and basic research. The high specificity of monoclonals found its
jection of the selected hybridoma, followed by regular aspiration of way in a number of laboratory applications. Many of the immunore-
the resulting immunoglobulin-­rich ascites produced. agents already used in nephelemetry, turbidimetry, ELISA or multi-
The spread was what would now be qualified as “viral.” Hundreds plex chemiluminescence were successfully replaced by monoclonals.
of abstracts and papers blossomed. In the early 1980s, Laurence Even in more fundamental work, Western blots found an increased
Boumsell and Alain Bernard, who had close relationships both with resolution by the use of monoclonals. The dissection of antigenic
Cesar Milstein and Jean Dausset, devised a way to bring some order epitopes became feasible.
to the ongoing folly. Rich of the experience of HLA workshops In the initial applications of immunophenotyping or immu-
heralded by their mentor Dausset, they organized a worldwide nochemistry, indirect methods were used, with anti-­mouse labeled
co-­operation under the name of Human Leukocyte Differentiation antisera as secondary layer. The reagent industry soon grabbed
Antigen (HLDA) workshops. 22 Indeed, most of the laboratories that the interest of providing purified monoclonal antibody solutions. It
had been investing in this new wave had been trying to use these must be acknowledged that the Ortho company was pioneer in this
new fantastic tools to find a better definition of white blood cells. It field. 28 Very early in the 1980s the antibodies produced by Patrice
must be reminded that, at that time, morphology, some enzymatic Kung were exploited and commercialized by this manufacturer as
reactions, and the use of polyclonal animal antisera were the only “OKT” or “Ortho Kung T”-­antibodies. OKT3, OKT4, and OKT8 were
means to (poorly) decipher leukocyte subsets. It was possible to certainly the first used in many laboratories to discover the exquisite
identify immunoglobulins on the surface of mature B cells and in the homeostasis of peripheral T-­cell subsets, initially only suspected by
cytoplasm of preB cells. 23,24 T-­lymphocytes, characterized after the the use of the abovementioned sophisticated rosette experiments.
work of John Miller on the thymus, 25 were tackled by sophisticated Quickly, fluorochrome-­
conjugated monoclonals became available
variations of their ability to collect sheep red blood cells around and greatly helped the development of flow cytometry.
them, the rosetting phenomenon. In this specific field of flow cytometry, the first steps were to
Human Leukocyte Differentiation Antigen workshops met with a move from indirect immunofluorescence toward direct labeling. The
tremendous success right from the start when the first proceedings all-­time winner, already developed in the late 70s for fluorescence
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microscopy, is indeed fluorescein isothiocyanate or FITC. This rather 6 | FRO M R E AG E NT S TO TH E R A PY,

sturdy fluorochrome is still much used but the collection of possible M O N O C LO N A L A NTI B O D I E S
fluorochromes is an ever-­growing field. Many of these substances BIOENGINEERING
are derived from algae or phytoplankton and are exquisitely sensitive
to light. This is why they are stored and commercialized in light-­proof Cesar Milstein is reported to have, very early upon the discovery of
vials and should be exposed to light for the smallest possible time monoclonal antibodies, envisioned that these manufactured immu-
before analyzing the cells or substances that have been expected noglobulins had a potential as drugs.
to be identified by immunofluorescence. This sensitivity to light With the appearance of monoclonal antibodies, two main chal-
and pH conditions is exacerbated in the more sophisticated fluoro- lenges faced this new era of immunotherapy: (a) obtain enough im-
chromes known as tandems. In this case, two molecules are bound munoglobulins for human applications, (b) avoid xeno-­immunization
together so that the fluorescence emitted by the first by excitation against mouse immunoglobulins. Furthermore, better knowledge of
at a given wavelength corresponds to the excitation wavelength of the mode of action of these new drugs led to attempts at improving
the second. This allows to use one laser source (that exciting the first the respective immune reactions initiated and improve the pharma-
fluorochrome) to trigger fluorochromes outside of the instrument's cokinetics of these new drugs.
wavelength, that is obtain light that could otherwise not be emitted. Several models were devised to engineer monoclonal antibodies
Besides these chemicals, pigments or proteins, another range of flu- in sufficient amounts, moving from the initial murine ascites
orescent probes is that of the Quantum dyes which are inorganic model.33,34 Some transgenic animals were developed. The eggs of
nanocrystals. More recently, in the sophisticated methods of mass transgenic hens and the milk of transgenic rabbits or goats have
cytometry, monoclonal antibody labels have been changed to rare been successfully used. Bacteria can be used, yet the glycosylation
earth metals (elemental isotopes). Detection of their presence on and assembly of immunoglobulins is not optimal in such cells and
labeled cells relies on mass spectrometry after nebulization of each transfectants may be too large for them. Non-mammalian insect or
cell in a plasma torch, thus assessing their “time of flight,” hence the plant cells were also transfected with various constructions, yielding
name of CyTOF for this methodology. 29 functional antibodies. The largest field, however, has been that of
mammalian cell lines. COS1 cells have been widely used for the rapid
production of a few micrograms per mL of antibodies, allowing for
5 | U N D E R S TA N D I N G TH E M O LEC U L A R quick screening.35 Mouse and human cell lines have also been tested.
BA S E S O F I M M U N O G LO B U LI N S D I V E R S IT Y However, Chinese hamster ovary cells (CHO) have become the sup-
port of choice. They allow for the production of high levels of anti-
In the late 1970s, though, it was still an unsolved puzzle how the bodies, up to tens or hundreds of milligrams per million cells per
specific DNA encoding mRNA for full immunoglobulins and the rele- day.35 These cells can grow at high densities in suspension with
vant proteins could fit the genome. There was a terrible size problem serum-­free medium of defined chemical composition. They produce
because according to the expected diversity of immunoglobulins, it mostly properly folded and glycosylated proteins and have been
was just impossible that each was encoded by a full gene. This would used for the production of most of the commercialized therapeutic
have far exceeded the 6 pg of DNA in each cell of a human body, not monoclonal antibodies. Some human cell lines have also been devel-
mentioning other species. The merit (and Nobel prize) goes to the oped for the transfection of constructs that will yield totally compat-
imaginative brain of Susumu Tonegawa, also working at the Basel ible glycoproteins.33
Institute, based on the fact, recognized by the mid-­60s, that the NH2 Immunogenicity was the reason why the first FDA approved
terminal domain of immunoglobulin chains was the most variable therapeutic monoclonal, OKT3, used in extreme conditions of
one. The idea that this strand of DNA, for each B-­lymphocyte, re- transplant rejection, did not really remain a valid solution. 36
sulted from the juxtaposition of small multiple segments had already The strategies used to reduce the immunogenicity of therapeu-
crossed the mind of Cesar Milstein. Tonegawa identified the fact tic monoclonal antibodies led to the current nomenclature used
that “repertoires” of small gene segments was what solved the rid- (Figure 1, Table 2). From fully murine immunoglobulins, genetic
dle.30 These repertoires are now fully mapped,31 for immunoglobulin engineering moved to chimeric antibodies retaining only the
heavy chains on chromosome 14, and for light chains on chromo- mouse Fab. This was the case for the successful anti-­CD20 ritux-
somes 2 and 22. “Rearranged” heavy chain genes each use one vari- imab, a chimeric IgG1 approved for non-­H odgkin's lymphoma in
34,37
able (V), one diversity (D), and one junction (J) segments, randomly 1997, representing the first mAb with oncologic indication
selected during B-­cell maturation in the bone marrow. Different V and the third approved therapeutic monoclonal. Monoclonal an-
and J segments also exist for light chains. The possible variations tibodies were then humanized and finally became fully human
combining the diversity of heavy chains and kappa or lambda light antibodies, the latter only retaining the most affine CDR (comple-
chains provide enough diversity to comply with the antigenic diver- mentary determining regions) amino acids within a wholly human
sity on our planet! Of note, the same puzzling phenomenon occurs immunoglobulin. Progress in genetic engineering and precise
early in the development of T cells in the thymus, with rearrange-
ments of coding segments from other T-­specific repertoires.32 1
Cells of simian (CV-­1) Origin with SV40.
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12       BÉNÉ

F I G U R E   1   Nomenclature of therapeutic monoclonal antibodies. “Momabs” are completely murine (and highly immunogenic).
Bioengineered antibodies have been devised to limit murine sequences to the antigen-­binding regions. Their various names thus depend
on the variable region of the immunoglobulin. The latter is fully murine in “ximabs”. The difference between humanized (-­zu) and fully
human monoclonals (-­u) depends on how close to a human sequence is the antigen-­binding structure. To produce fully human monoclonals,
the mouse genome is modified to harbor the human immunoglobulin locus before immunization 51,52 [Colour figure can be viewed at
wileyonlinelibrary.com]

TA B L E   2   Examples of different types of therapeutic antibodies for the treatment of hematological malignancies, with decreasing
antigenicity. The isotype used is important for the half-­life of the product and to favor or alleviate interactions with Fc receptors

Name Type Target Isotype Function Mechanism

OKT3 Murine CD3 IgGa2 Cytotoxic

Rituximab Chimeric CD20 IgG1 Cytotoxic, proapoptotic ADCC, CDC
Obinutuzumab Humanized CD20 IgG1 Cytotoxic, glycosylation favors ADCC, ADCP
FcR interactions
Ofatumumab Fully human CD20 IgG1 Cytotoxic, proapoptotic ADCC, CDC
Nivolumab Fully human PD-­1 IgG4 Antagonist Blocks PDL-­1
signaling

deciphering of antibody-coding regions allowed for a variety of have been devised to modulate these interactions and impact on the
developments to improve the specificity and affinity of therapeu- drug's pharmacokinetics.39 Another strategy is to generate hybrid
tic recombinant monoclonal antibodies. Such strategies also were IgG2-­CH1 (which does not bind complement) and IgG4-­CH3-­CG3
applied to improve the pharmacokinetics of these new drugs, for (which do not bind FcR), preferred for antagonist or agonist anti-
instance favoring their recycling by interaction with the neonatal bodies that should not lead to cytotoxicity. This was applied to the
Fcn receptor. 38 generation of eculizumab, the anti-­C5 antibody stabilizing patients
Currently, most of therapeutic monoclonals are thus human with paroxysmal nocturnal hemoglobinuria.40 Indeed, besides cy-
IgG1,34 which recognize the chosen target by their Fab portion totoxicity to remove tumor cells or adverse immune reactions, re-
and display effector functions through their Fc. The latter include cent remarkable applications involve manipulation of the immune
complement binding in complement depending cytotoxicity (CDC) system. The development of immune checkpoint inhibitors reviving
and attachment to Fc receptors (FcR) on phagocytes in antibody-­ the activity of anergic or exhausted T cells has led to a whole field
dependent cellular phagocytosis (ADCP) and on natural killer cells of antitumoral strategies based on reactivation of the patient's own
in antibody-­
dependent cellular cytotoxicity (ADCC). These func- defense systems.41
tions are mostly used for therapeutic preparations directed to Other approaches removed the Fc fragment when the latter was
tumor cells such as lymphoma cells. Variations in Fc glycosylation not necessary. In 1994, abciximab, a chimeric anti-­GPIIb/IIIa Fab

F I G U R E   2   A brief history of
immunoglobulins and monoclonal
antibodies [Colour figure can be viewed at
wileyonlinelibrary.com]
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fragment inhibiting platelet aggregation in cardiovascular diseases, 7 | CO N C LU S I O N S

was approved by the FDA, representing the second therapeutic
mAb.42 The unique properties of immunoglobulins Fc fragments It would be an amazing scenario to see Metchnikoff, Ehrlich, von
have also been used to produce chimeric drugs combining them Behring or Kitasato return to our world and see what happened fol-
with, for instance, soluble cytokine receptors. The family of anti-­TNF lowing their initial discoveries (Figure 2). That they were right on so
bioactives has thus completely revolutionized the treatment of such many points of their observations certainly paved the way for other
inflammatory diseases as rheumatoid arthritis. The Fc fragment of daring and imaginative researchers to follow their path. It would
IgG also increases the half-­life of recombinant proteins such as EPO probably be even more puzzling to show the pioneers of vaccination
or coagulation factors.43 who are ancient Chinese, the “younger” Jenner or Pasteur, the mo-
The medical field of application of these immunotherapies is in- lecular intricacies, and beautiful mechanics underlying their intuition
creasing in range and involves hemato-­oncology, solid tumors, he- that the exquisite specificity of the immune system can be manipu-
mophilia, coagulation, asthma, hypercholesterolemia, age-­
related lated for the benefits of health.
macular degeneration, chronic inflammatory diseases, diabetes…to
cite a few.33-35
C O N FL I C T O F I N T E R E S T
Therapeutic monoclonal antibodies may act as “nude” mole-
cules, acting by themselves, or as “magic bullets,” vectorizing drugs The author has no competing interests.
toward specific targets. These immunotoxins have been developed
in a number of applications, essentially in oncology.44 Regular in-
ORCID
sights into this still fully growing fields are provided by the series
“Antibodies to watch,” the issue celebrating its tenth year providing Marie C Béné  https://orcid.org/0000-0002-6569-7414
an even broader view than usual.45
More recently, an idea that had emerged as early as 1960 46,47
but was not really pursued until 199548 has reached the level of drug REFERENCES

development, namely bispecific antibodies. In this type of construc- 1. von Behring E, Kitasato S. Über das Zustandekommen der
tion, two variable domains of different specificity are combined in Diphtherie-­ Immunität und der Tetanus-­ Immunität bei Thieren.
Dtsch Med Wschr. 1890;16:1113‐1114.
order to connect two different types of cells. The most successful to
2. Lindenmann J. Origin of the terms ‘antibody’ and ‘antigen’. Scand J
date bring T cells, through their anti-CD3 moiety, toward malignant Immunol. 1984;19:281‐285.
B cells expressing CD19.49 More constructions are in progress for 3. Nicolle M. Une conception générale des anticorps et de leurs effets.
other onco-­hematologic diseases or other cancers. CR Soc Biol (Paris). 1907;63:77.
4. Deutsch L. Contribution à I’étude de l'origine des anticorps typh-
Finally, following a long series of bioengineering strategies, the
iques. Ann Inst Pasteur. 1899;13:689.
unique ability of immunoglobulins Fab to recognize antigens directly
5. Tizzoni G, Cattani G. Fernere Untersuchungen liber das tetanus-­
has been put to use to improve T-­cell targeting and activation in fully antitoxin. Zentralbl Bakteriol Mikrobiol Hyg [A]. 1891;10:33.
bioengineered Chimeric Antigen Receptor T cells or CAR T cells. 50 6. Tiselius A. Electrophoresis of serum globulin I. Biochem J.
The Fab portion of an immunoglobulin is modified to produce a sin- 1937;31:313-317.
7. Edelman GM. Dissociation of y-­ globulin. J Am chem Soc.
gle chain variable fragment (scFv) associating the variable domains
1959;81:3155.
of the heavy and light chain. Genes coding for this construction are 8. Ceppellini R, Dray S, Edelman G, et al. Nomenclature for human
then associated to those encoding a transmembrane portion and in- immunoglobulins. Immunochemistry. 1964;1:145-149.
tracytoplasmic T-cell activation domains. The first CAR T-cells only 9. Heremans JF. Les Globulines Sériques du Système Gamma. Brussels:
Editions Arscia; 1960.
associated the CD3 zeta chain of the T-­cell receptor. Nowadays, CAR
10. Ehrlich P. Die Seitenkettentheorie und ihre Gegner. Münchner Med
T cells of three different generations have been produced, associat- Wochenschr 1901;2123‐2124.
ing cytoplasmic portions of other co-­activation molecules such as 11. Valent P, Groner B, Schumacher U, et al. Paul Ehrlich (1854-­1915)
CD28 and/or 4-1BB. This expanding field triggers increasingly imag- and His Contributions to the Foundation and Birth of Translational
Medicine. J Innate Immun. 2016;8:111-120.
inative solutions, based on the combination of the now well-­known
12. Cohn EJ, Oncley JL, Strong LE, Hughes WL Jr, Armstrong SH Jr.
molecular portions of immunoglobulins, T-­cell receptors and co-­ Chemical, clinical, and immunological studies on the products of
activation signals in a genial genetic meccano. Modified T cells from human plasma fractionation. I. The characterization of the protein
the patient are currently used, reinfused after a few days or weeks. fractions of human plasma. J Clin Invest. 1944;23:417‐432.
13. Imbach P, Barandun S, Baumgartner C, Hirt A, Hofer F, Wagner HP.
“Off the shelf” allogeneic CAR T-cells from donors, modified not only
High-­dose intravenous gammaglobulin therapy of refractory, in
for their CAR portion but also to remove possible alloreactivity are particular idiopathic, thrombocytopenia in childhood. Helv Paediatr
also developed. Some models even include suicide genes allowing to Acta. 1981;46:81-86.
induce the destruction of the infused CAR T-cells, should they yield 14. Hooper JA. The history and evolution of immunoglobulin products
and their clinical indications. LymphoSign J. 2015;2:181-194.
adverse reactions.
|
14       BÉNÉ

15. Porter RR. The structure of y-globulin and antibodies. In: Gellhorn 35. Trill JJ, Shatzman AR, Ganguly S. Production of monoclonal antibod-
A, Hirschberg E, ed. Basic Problems of Neoplastic Disease. New York, ies in COS and CHO cells. Curr Opin Biotechnol. 1995;6(553):560.
NY: Columbia University Press;1962:177. 36. Sgro C. Side-­effects of a monoclonal antibody, muromonab CD3/or-
16. Jones HB. Papers on chemical pathology; prefaced by the thoclone OKT3: a bibliographic review. Toxicology. 1995;105:23e9.
Gulstonian lectures read at the Royal College of Physicians. Lecture 37. Marcus R, Hagenbeek A. The therapeutic use of rituximab in non-­
3. Lancet. 1846;2: 88‐92. Hodgkin's lymphoma. Eur J Haematol Suppl. 2007;67(5):14.
17. Korngold L, Lipari R. Multiple myeloma proteins. III. The antigenic 38. Sheridan D, Yu ZX, Zhang Y, et al. Design and preclinical charac-
relationship of Bence-­Jones proteins to normal y-­globulin and mul- terization of ALXN1210: a novel anti-­C5 antibody with extended
tiple myeloma serum proteins. Cancer. 1956;9:262. duration of action. PLoS ONE. 2018;13:e0195909.
18. Gowans JL, McGregor DD, Cowan DM, Ford CE. Initiation of Immune 39. Jefferis R. Posttranslational modifications and the immunogenicity
Responses by Small Lymphocytes. Nature. 1962;196:651-652. of biotherapeutics. J Immunol Res. 2016;2016:5358272.
19. Jerne NK, Henry C, Nordin AA, Fuji H, Koros AM, Lefkovits I. 4 0. Rother RP, Rollins SA, Mojcik CF, Brodsky RA, Bell L. Discovery
Plaque forming cells: methodology and theory. Transplant Rev. and development of the complement inhibitor eculizumab for the
1974;18:130-191. treatment of paroxysmal nocturnal hemoglobinuria. Nat Biotechnol.
20. Halaby DM, Mornon JP. The immunoglobulin superfamily: an in- 2007;25(1256):1264.
sight on its tissular, species, and functional diversity. J Mol Evol. 41. Rotte A, Jin JY, Lemaire V. Mechanistic overview of immune check-
1998;46:389-400. points to support the rational design of their combinations in cancer
21. Köhler G, Milstein C. Continuous cultures of fused cells secreting immunotherapy. Ann Oncol. 2018;29(71):83.
antibody of predefined specificity. Nature. 1975;256:495-497. 42. Foster RH, Wiseman LR. Abciximab. An updated review of its use in
22. Bernard A, Boumsell L. The clusters of differentiation (CD) de- ischaemic heart disease. Drugs 1998;56:629‐665.
fined by the First International Workshop on Human Leucocyte 43. Kontermann RE. Strategies to extend plasma half-­lives of recombi-
Differentiation Antigens. Hum Immunol. 1984;11:1-10. nant antibodies. BioDrugs. 2009;23(93):109.
23. Fröland S, Natvig JB, Berdal P. Surface-­ bound immunoglobu- 4 4. Wayne AS, Fitzgerald DJ, Kreitman RJ, Pastan I. Immunotoxins for
lin as a marker of B lymphocytes in man. Nat New Biol. 1971 Dec leukemia. Blood 2014;123:2470‐2477.
22;234(51):251-252. 45. Kaplon H, Reichert JM. Antibodies to watch in 2019. MAbs.
24. Guglielmi P, Preud'homme JL. Immunoglobulin expression in human 2019;11:219‐238.
lymphoblastoid cell lines with early B cell features. Scand J Immunol. 46. Nisonoff A, Rivers MM. Recombination of a mixture of univalent
1981;13:303‐311. antibody fragments of different specificity. Arch Biochem Biophys.
25. Miller JF. Role of the cells which originate from the thymus and 1961;93:460‐462.
bone marrow. Ann Immunol (Paris). 1974;125C:213-229. 47. Brinkmann U, Kontermann RE. The making of bispecific antibodies.
26. Engel P, Boumsell L, Balderas R, et al. CD Nomenclature 2015: MAbs. 2017;9:182‐212.
human Leukocyte Differentiation Antigen Workshops as a Driving 48. Davico Bonino L, De Monte LB, Spagnoli GC, et al. Bispecific mono-
Force in Immunology. J Immunol. 2015;195:4555-4563. clonal antibody anti-­CD3 x anti-­tenascin: an immunotherapeutic
27. Zola H, Swart B. The human leucocyte differentiation antigens agent for human glioma. Int J Cancer 1995;61:509‐515.
(HLDA) workshops: the evolving role of antibodies in research, di- 49. Smits NC, Sentman CL. Bispecific T-­Cell engagers (BiTEs) as treat-
agnosis and therapy. Cell Res. 2005;15:691-698. ment of B-­Cell lymphoma. J Clin Oncol 2016;34:1131‐1133.
28. Reinherz EL, Kung PC, Goldstein G, Schlossman SF. Further charac- 50. Levine BL, Miskin J, Wonnacott K, Keir C. Global Manufacturing of
terization of the human inducer T cell subset defined by monoclonal CAR T Cell Therapy. Mol Ther Methods Clin Dev 2017;4:92‐101.
antibody. J Immunol. 1979;123:2894‐2896. 51. Jones TD, Carter PJ, Plückthun A, et al. The INNs and outs of anti-
29. Nolan GP. Flow cytometry in the post fluorescence era. Best Pract body nonproprietary names. MAbs. 2016;8:1.9
Res Clin Haematol. 2011;24:505-508. 52. Mallbris L, Davies J, Glasebrook A, Tang Y, Glaesner W, Nickoloff
3 0. Tonegawa S. Somatic generation of antibody diversity. Nature. BJ. Molecular insights into fully human and humanized monoclo-
1983;302:575-581. nal antibodies: what are the differences and should dermatologists
31. Lefranc MP. Antibody Informatics: IMGT, the International care? J Clin Aesthet Dermatol. 2016;9:13‐15.
ImMunoGeneTics Information System. Microbiol Spectr. 2014;Apr
2(2).
32. Davis MM. Molecular genetics of the T cell-­receptor beta chain.
How to cite this article: Béné MC. The wonderful story of
Annu Rev Immunol. 1985;3(537):560.
monoclonal antibodies. Int J Lab Hematol.
33. Lalonde ME, Durocher Y. Therapeutic glycoprotein production in
mammalian cells. J Biotechnol. 2017;251:128‐240. 2019;41(Suppl. 1):8‐14. https://doi.org/10.1111/ijlh.13006
3 4. Dos Santos ML, Quintilio W, Manieri TM, Tsuruta LR, MoroBraz
AM. Advances and challenges in therapeutic monoclonal antibodies
drug development. Braz J Pharm Sci. 2018;54(Special):e01007.