Abstract
Monoclonal antibodies are among the most significant biological tools used in medicine and biology that have revolutionized the field of diagnostics, therapeutics, and targeted drug delivery systems for many diseases. Among them, rabbit monoclonal antibodies have attracted significant attention for having high affinity and specificity. During the past few decades, different techniques have been developed to produce monoclonal antibodies. Single B cell cloning technology offers many advantages compared to other methods and has been used to generate monoclonal antibodies from different species including rabbits. This review briefly describes some of these methods, with main focus on single B cell cloning and production of rabbit monoclonal antibodies.
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References
Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497
Tansey EM, Catterall PP (1994) Monoclonal antibodies: a witness seminar in contemporary medical history. Med Hist 38:322–327
Zhang H, Chen J (2018) Current status and future directions of cancer immunotherapy. J Cancer 9:1773–1781
Henricks LM, Schellens JH, Huitema AD et al (2015) The use of combinations of monoclonal antibodies in clinical oncology. Cancer Treat Rev 41:859–867
Siddiqui MZ (2010) Monoclonal antibodies as diagnostics; an appraisal. Indian J Pharm Sci 72:12–17
Chan AC, Carter PJ (2010) Therapeutic antibodies for autoimmunity and inflammation. Nat Rev Immunol 10:301–316
Scott AM, Wolchok JD, Old LJ (2012) Antibody therapy of cancer. Nat Rev Cancer 12:278–287
Reichert JM (2017) Antibodies to watch in 2017. MAbs 9:167–181
Kaplon H, Reichert JM (2018) Antibodies to watch in 2018. MAbs 10:183–203
Rossi S, Laurino L, Furlanetto A et al (2005) Rabbit monoclonal antibodies: a comparative study between a novel category of immunoreagents and the corresponding mouse monoclonal antibodies. Am J Clin Pathol 124:295–302
Weber J, Peng H, Rader C (2017) From rabbit antibody repertoires to rabbit monoclonal antibodies. Exp Mol Med 49:e305. https://doi.org/10.1038/emm.2017.23
Landry JP, Ke Y, Yu GL et al (2015) Measuring affinity constants of 1450 monoclonal antibodies to peptide targets with a microarray-based label-free assay platform. J Immunol Methods 417:86–96
Shawler DL, Bartholomew RM, Smith LM et al (1985) Human immune response to multiple injections of murine monoclonal IgG. J Immunol 135:1530–1535
Rader C, Ritter G, Nathan S et al (2000) The rabbit antibody repertoire as a novel source for the generation of therapeutic human antibodies. J Biol Chem 275:13668–13676
Smith GP (1985) Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 228:1315–1317
Murai T, Ueda M, Yamamura M et al (1997) Construction of a starch-utilizing yeast by cell surface engineering. Appl Environ Microbiol 63:1362–1366
Rakestraw JA, Aird D, Aha PM et al (2011) Secretion-and-capture cell-surface display for selection of target-binding proteins. Protein Eng Des Sel 24:525–530
Roberts RW, Szostak JW (1997) RNA-peptide fusions for the in vitro selection of peptides and proteins. Proc Natl Acad Sci U S A 94:12297–12302
Spieker-Polet H, Sethupathi P, Yam PC et al (1995) Rabbit monoclonal antibodies: generating a fusion partner to produce rabbit-rabbit hybridomas. Proc Natl Acad Sci U S A 92:9348–9352
Yam PC, Knight KL (2014) Generation of rabbit monoclonal antibodies. Methods Mol Biol 1131:71–79
Tiller T (2011) Single B cell antibody technologies. New Biotechnol 28:453–457
Smith K, Garman L, Wrammert J et al (2009) Rapid generation of fully human monoclonal antibodies specific to a vaccinating antigen. Nat Protoc 4:372–384
Clargo AM, Hudson AR, Ndlovu W et al (2014) The rapid generation of recombinant functional monoclonal antibodies from individual, antigen-specific bone marrow-derived plasma cells isolated using a novel fluorescence-based method. MAbs 6:143–159
Holzlohner P, Hanack K (2017) Generation of murine monoclonal antibodies by hybridoma technology. J Vis Exp 119. https://doi.org/10.3791/54832
Raybould TJ, Takahashi M (1988) Production of stable rabbit-mouse hybridomas that secrete rabbit mAb of defined specificity. Science 240:1788–1790
Yarmush ML, Gates FT 3rd, Weisfogel DR et al (1980) Identification and characterization of rabbit-mouse hybridomas secreting rabbit immunoglobulin chains. Proc Natl Acad Sci U S A 77:2899–2903
Dreher K, Sogn JA, Gates FT 3rd et al (1983) Allotype-defined mRNA for rabbit immunoglobulin H and L chains isolated from rabbit-mouse hybridomas. J Immunol 130:442–448
Pytela R, Zhu W, Ke Y et al (2005) Fusion partner for production of monoclonal rabbit antibodies US Patent US7429487B2
Dessain SK, Adekar SP, Stevens JB et al (2004) High efficiency creation of human monoclonal antibody-producing hybridomas. J Immunol Methods 291:109–122
Li J, Sai T, Berger M et al (2006) Human antibodies for immunotherapy development generated via a human B cell hybridoma technology. Proc Natl Acad Sci U S A 103:3557–3562
Breitling F, Dubel S, Seehaus T et al (1991) A surface expression vector for antibody screening. Gene 104:147–153
McCafferty J, Griffiths AD, Winter G et al (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554
Barbas CF 3rd, Kang AS, Lerner RA et al (1991) Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc Natl Acad Sci U S A 88:7978–7982
Hammers CM, Stanley JR (2014) Antibody phage display: technique and applications. J Invest Dermatol 134:1–5
Pande J, Szewczyk MM, Grover AK (2010) Phage display: concept, innovations, applications and future. Biotechnol Adv 28:849–858
Sidhu SS, Li B, Chen Y et al (2004) Phage-displayed antibody libraries of synthetic heavy chain complementarity determining regions. J Mol Biol 338:299–310
Rakonjac J, Russel M, Khanum S et al (2017) Filamentous phage: structure and biology. Adv Exp Med Biol 1053:1–20
Ledsgaard L, Kilstrup M, Karatt-Vellatt A et al (2018) Basics of antibody phage display technology. Toxins 10:236. https://doi.org/10.3390/toxins10060236
Carmen S, Jermutus L (2002) Concepts in antibody phage display. Brief Funct Genomic Proteomic 1:189–203
Frenzel A, Kugler J, Helmsing S et al (2017) Designing human antibodies by phage display. Transfus Med Hemother 44:312–318
Moghaddam A, Borgen T, Stacy J et al (2003) Identification of scFv antibody fragments that specifically recognise the heroin metabolite 6-monoacetylmorphine but not morphine. J Immunol Methods 280:139–155
Blokzijl A, Zieba A, Hust M et al (2016) Single chain antibodies as tools to study transforming growth factor-beta-regulated SMAD proteins in proximity ligation-based pharmacological screens. Mol Cell Proteomics 15:1848–1856
Zhang Z, Liu H, Guan Q et al (2017) Advances in the isolation of specific monoclonal rabbit antibodies. Front Immunol 8:494. https://doi.org/10.3389/fimmu
Starkie DO, Compson JE, Rapecki S et al (2016) Generation of recombinant monoclonal antibodies from immunised mice and rabbits via flow cytometry and sorting of antigen-specific IgG+ memory B cells. PLoS One 11:e0152282. https://doi.org/10.1371/journal.pone.0152282
Pattengale PK, Smith RW, Gerber P (1973) Selective transformation of B lymphocytes by E.B. virus. Lancet 2:93–94
Kozbor D, Roder JC (1981) Requirements for the establishment of high-titered human monoclonal antibodies against tetanus toxoid using the Epstein-Barr virus technique. J Immunol 127:1275–1280
Beerli RR, Rader C (2010) Mining human antibody repertoires. MAbs 2:365–378
Shammah S, Mantovani TL, Dalla-Favera R et al (1993) Generation of human monoclonal antibodies by transformation of lymphoblastoid B cells with ras oncogene. J Immunol Methods 160:19–25
Rashidian J, Copaciu R, Su Q et al (2017) Generation and performance of R132H mutant IDH1 rabbit monoclonal antibody. Antibodies 6. https://doi.org/10.3390/antib6040022
Ouisse LH, Gautreau-Rolland L, Devilder MC et al (2017) Antigen-specific single B cell sorting and expression-cloning from immunoglobulin humanized rats: a rapid and versatile method for the generation of high affinity and discriminative human monoclonal antibodies. BMC Biotechnol 17:3. https://doi.org/10.1186/s12896-016-0322-5
Kuppers R, Zhao M, Hansmann ML et al (1993) Tracing B cell development in human germinal centres by molecular analysis of single cells picked from histological sections. EMBO J 12:4955–4967
Tiller T, Meffre E, Yurasov S et al (2008) Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. J Immunol Methods 329:112–124
Lagerkvist AC, Furebring C, Borrebaeck CA (1995) Single, antigen-specific B cells used to generate Fab fragments using CD40-mediated amplification or direct PCR cloning. BioTechniques 18:862–869
Jin A, Ozawa T, Tajiri K et al (2011) A Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array. Nat Protoc 6:668–676
Park S, Han J, Kim W et al (2011) Rapid selection of single cells with high antibody production rates by microwell array. J Biotechnol 156:197–202
Meijer PJ, Andersen PS, Haahr Hansen M et al (2006) Isolation of human antibody repertoires with preservation of the natural heavy and light chain pairing. J Mol Biol 358:764–772
Franz B, May KF Jr, Dranoff G et al (2011) Ex vivo characterization and isolation of rare memory B cells with antigen tetramers. Blood 118:348–357
Amanna IJ, Slifka MK (2006) Quantitation of rare memory B cell populations by two independent and complementary approaches. J Immunol Methods 317:175–185
Doucett VP, Gerhard W, Owler K et al (2005) Enumeration and characterization of virus-specific B cells by multicolor flow cytometry. J Immunol Methods 303:40–52
Di Niro R, Mesin L, Raki M et al (2010) Rapid generation of rotavirus-specific human monoclonal antibodies from small-intestinal mucosa. J Immunol 185:5377–5383
Tiller T, Busse CE, Wardemann H (2009) Cloning and expression of murine Ig genes from single B cells. J Immunol Methods 350:183–193
Kurosawa N, Yoshioka M, Fujimoto R et al (2012) Rapid production of antigen-specific monoclonal antibodies from a variety of animals. BMC Biol 10:80. https://doi.org/10.1186/1741-7007-10-80
Seeber S, Ros F, Thorey I et al (2014) A robust high throughput platform to generate functional recombinant monoclonal antibodies using rabbit B cells from peripheral blood. PLoS One 9:e86184. https://doi.org/10.1371/journal.pone.0086184
Ozawa T, Kishi H, Muraguchi A (2006) Amplification and analysis of cDNA generated from a single cell by 5′-RACE: application to isolation of antibody heavy and light chain variable gene sequences from single B cells. BioTechniques 40:469–470
Liao HX, Levesque MC, Nagel A et al (2009) High-throughput isolation of immunoglobulin genes from single human B cells and expression as monoclonal antibodies. J Virol Methods 158:171–179
Ojima-Kato T, Hashimura D, Kojima T et al (2015) In vitro generation of rabbit anti-Listeria monocytogenes monoclonal antibody using single cell based RT-PCR linked cell-free expression systems. J Immunol Methods 427:58–65
Kivi G, Teesalu K, Parik J et al (2016) HybriFree: a robust and rapid method for the development of monoclonal antibodies from different host species. BMC Biotechnol 16:2. https://doi.org/10.1186/s12896-016-0232-6
Gray SA, Moore M, VandenEkart EJ et al (2016) Selection of therapeutic H5N1 monoclonal antibodies following IgVH repertoire analysis in mice. Antivir Res 131:100–108
Reddy ST, Ge X, Miklos AE et al (2010) Monoclonal antibodies isolated without screening by analyzing the variable-gene repertoire of plasma cells. Nat Biotechnol 28:965–969
Saggy I, Wine Y, Shefet-Carasso L et al (2012) Antibody isolation from immunized animals: comparison of phage display and antibody discovery via V gene repertoire mining. Protein Eng Des Sel 25:539–549
Wang B, Kluwe CA, Lungu OI et al (2015) Facile discovery of a diverse panel of anti-Ebola virus antibodies by immune repertoire mining. Sci Rep 5:13926. https://doi.org/10.1038/srep13926
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Rashidian, J., Lloyd, J. (2020). Single B Cell Cloning and Production of Rabbit Monoclonal Antibodies. In: Zielonka, S., Krah, S. (eds) Genotype Phenotype Coupling. Methods in Molecular Biology, vol 2070. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9853-1_23
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DOI: https://doi.org/10.1007/978-1-4939-9853-1_23
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