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Synthetic approach to the generation of antibody diversity
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  • Journal title : BMB Reports
  • Volume 48, Issue 9,  2015, pp.489-494
  • Publisher : Korean Society for Biochemistry and Molecular Biology
  • DOI : 10.5483/BMBRep.2015.48.9.120
 Title & Authors
Synthetic approach to the generation of antibody diversity
Shim, Hyunbo;
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The in vitro antibody discovery technologies revolutionized the generation of target-specific antibodies that traditionally relied on the humoral response of immunized animals. An antibody library, a large collection of diverse, pre-constructed antibodies, can be rapidly screened using in vitro display technologies such as phage display. One of the keys to successful in vitro antibody discovery is the quality of the library diversity. Antibody diversity can be obtained either from natural B-cell sources or by the synthetic methods that combinatorially generate random nucleotide sequences. While the functionality of a natural antibody library depends largely upon the library size, various other factors can affect the quality of a synthetic antibody library, making the design and construction of synthetic antibody libraries complicated and challenging. In this review, we present various library designs and diversification methods for synthetic antibody library. From simple degenerate oligonucleotide synthesis to trinucleotide synthesis to physicochemically optimized library design, the synthetic approach is evolving beyond the simple emulation of natural antibodies, into a highly sophisticated method that is capable of producing high quality antibodies suitable for therapeutic, diagnostic, and other demanding applications. [BMB Reports 2015; 48(9): 489-494]
Antibody library;Phage display;Synthetic antibody diversity;
 Cited by
Genetically modified bacteriophages, Integr. Biol., 2016, 8, 4, 465  crossref(new windwow)
Kohler G and Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495-497 crossref(new window)

Bradbury AR, Sidhu S, Dubel S and McCafferty J (2011) Beyond natural antibodies: the power of in vitro display technologies. Nat Biotechnol 29, 245-254 crossref(new window)

Sheets MD, Amersdorfer P, Finnern R et al (1998) Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens. Proc Natl Acad Sci U S A 95, 6157-6162 crossref(new window)

Loset GA, Lobersli I, Kavlie A et al (2005) Construction, evaluation and refinement of a large human antibody phage library based on the IgD and IgM variable gene repertoire. J Immunol Methods 299, 47-62 crossref(new window)

de Haard HJ, van Neer N, Reurs A et al (1999) A large non-immunized human Fab fragment phage library that permits rapid isolation and kinetic analysis of high affinity antibodies. J Biol Chem 274, 18218-18230 crossref(new window)

Marks JD, Hoogenboom HR, Bonnert TP, McCafferty J, Griffiths AD and Winter G (1991) By-passing immunization. Human antibodies from V-gene libraries displayed on phage. J Mol Biol 222, 581-597 crossref(new window)

Schwimmer LJ, Huang B, Giang H et al (2013) Discovery of diverse and functional antibodies from large human repertoire antibody libraries. J Immunol Methods 391, 60-71 crossref(new window)

Nissim A, Hoogenboom HR, Tomlinson IM et al (1994) Antibody fragments from a 'single pot' phage display library as immunochemical reagents. EMBO J 13, 692-698

Pini A, Viti F, Santucci A et al (1998) Design and use of a phage display library. Human antibodies with subnanomolar affinity against a marker of angiogenesis eluted from a two-dimensional gel. J Biol Chem 273, 21769-21776 crossref(new window)

Silacci M, Brack S, Schirru G et al (2005) Design, construction, and characterization of a large synthetic human antibody phage display library. Proteomics 5, 2340-2350 crossref(new window)

Yang HY, Kang KJ, Chung JE and Shim H (2009) Construction of a large synthetic human scFv library with six diversified CDRs and high functional diversity. Mol Cells 27, 225-235 crossref(new window)

Knappik A, Ge L, Honegger A et al (2000) Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides. J Mol Biol 296, 57-86 crossref(new window)

Rothe C, Urlinger S, Lohning C et al (2008) The human combinatorial antibody library HuCAL GOLD combines diversification of all six CDRs according to the natural immune system with a novel display method for efficient selection of high-affinity antibodies. J Mol Biol 376, 1182-1200 crossref(new window)

Prassler J, Thiel S, Pracht C et al (2011) HuCAL PLATINUM, a synthetic Fab library optimized for sequence diversity and superior performance in mammalian expression systems. J Mol Biol 413, 261-278 crossref(new window)

Tiller T, Schuster I, Deppe D et al (2013) A fully synthetic human Fab antibody library based on fixed VH/VL framework pairings with favorable biophysical properties. MAbs 5, 445-470 crossref(new window)

Sidhu SS and Fellouse FA (2006) Synthetic therapeutic antibodies. Nat Chem Biol 2, 682-688 crossref(new window)

Steinwand M, Droste P, Frenzel A, Hust M, Dubel S and Schirrmann T (2014) The influence of antibody fragment format on phage display based affinity maturation of IgG. MAbs 6, 204-218 crossref(new window)

Hudson PJ and Kortt AA (1999) High avidity scFv multimers; diabodies and triabodies. J Immunol Methods 231, 177-189 crossref(new window)

Desiderio A, Franconi R, Lopez M et al (2001) A semi-synthetic repertoire of intrinsically stable antibody fragments derived from a single-framework scaffold. J Mol Biol 310, 603-615 crossref(new window)

Rader C and Barbas CF 3rd (1997) Phage display of combinatorial antibody libraries. Curr Opin Biotechnol 8, 503-508 crossref(new window)

Kim YJ, Neelamegam R, Heo MA, Edwardraja S, Paik HJ and Lee SG (2008) Improving the productivity of single-chain Fv antibody against c-Met by rearranging the order of its variable domains. J Microbiol Biotechnol 18, 1186-1190

Luo D, Mah N, Krantz M et al (1995) Vl-linker-Vh ori- entation-dependent expression of single chain Fv-containing an engineered disulfide-stabilized bond in the framework regions. J Biochem 118, 825-831

Liu A, Ye Y, Chen W, Wang X and Chen F (2015) Expression of V(H)-linker-V(L) orientation-dependent single-chain Fv antibody fragment derived from hybridoma 2E6 against aflatoxin B1 in Escherichia coli. J Ind Microbiol Biotechnol 42, 255-262 crossref(new window)

Dolezal O, Pearce LA, Lawrence LJ, McCoy AJ, Hudson PJ and Kortt AA (2000) ScFv multimers of the anti-neuraminidase antibody NC10: shortening of the linker in single-chain Fv fragment assembled in V(L) to V(H) orientation drives the formation of dimers, trimers, tetramers and higher molecular mass multimers. Protein Eng 13, 565-574 crossref(new window)

Arnaout R, Lee W, Cahill P et al (2011) High-resolution description of antibody heavy-chain repertoires in humans. PLoS One 6, e22365 crossref(new window)

Soderlind E, Strandberg L, Jirholt P et al (2000) Recombining germline-derived CDR sequences for creating diverse single-framework antibody libraries. Nat Biotechnol 18, 852-856 crossref(new window)

Chothia C and Lesk AM (1987) Canonical structures for the hypervariable regions of immunoglobulins. J Mol Biol 196, 901-917 crossref(new window)

Al-Lazikani B, Lesk AM and Chothia C (1997) Standard conformations for the canonical structures of immunoglobulins. J Mol Biol 273, 927-948 crossref(new window)

Vargas-Madrazo E, Lara-Ochoa F and Almagro JC (1995) Canonical structure repertoire of the antigen-binding site of immunoglobulins suggests strong geometrical restrictions associated to the mechanism of immune recognition. J Mol Biol 254, 497-504 crossref(new window)

Van den Brulle J, Fischer M, Langmann T et al (2008) A novel solid phase technology for high-throughput gene synthesis. Biotechniques 45, 340-343 crossref(new window)

Zhai W, Glanville J, Fuhrmann M et al (2011) Synthetic antibodies designed on natural sequence landscapes. J Mol Biol 412, 55-71 crossref(new window)

Steipe B, Schiller B, Pluckthun A and Steinbacher S (1994) Sequence statistics reliably predict stabilizing mutations in a protein domain. J Mol Biol 240, 188-192 crossref(new window)

Lee CV, Liang WC, Dennis MS, Eigenbrot C, Sidhu SS and Fuh G (2004) High-affinity human antibodies from phage-displayed synthetic Fab libraries with a single framework scaffold. J Mol Biol 340, 1073-1093 crossref(new window)

MacCallum RM, Martin AC and Thornton JM (1996) Antibody-antigen interactions: contact analysis and binding site topography. J Mol Biol 262, 732-745 crossref(new window)

Fellouse FA, Wiesmann C and Sidhu SS (2004) Synthetic antibodies from a four-amino-acid code: a dominant role for tyrosine in antigen recognition. Proc Natl Acad Sci U S A 101, 12467-12472 crossref(new window)

Fellouse FA, Li B, Compaan DM, Peden AA, Hymowitz SG and Sidhu SS (2005) Molecular recognition by a binary code. J Mol Biol 348, 1153-1162 crossref(new window)

Fellouse FA, Esaki K, Birtalan S et al (2007) High-throughput generation of synthetic antibodies from highly functional minimalist phage-displayed libraries. J Mol Biol 373, 924-940 crossref(new window)

Persson H, Ye W, Wernimont A et al (2013) CDR-H3 diversity is not required for antigen recognition by synthetic antibodies. J Mol Biol 425, 803-811 crossref(new window)

Mahon CM, Lambert MA, Glanville J et al (2013) Comprehensive interrogation of a minimalist synthetic CDR-H3 library and its ability to generate antibodies with therapeutic potential. J Mol Biol 425, 1712-1730 crossref(new window)

Ewert S, Huber T, Honegger A and Pluckthun A (2003) Biophysical properties of human antibody variable domains. J Mol Biol 325, 531-553 crossref(new window)

Carton JM, Sauerwald T, Hawley-Nelson P et al (2007) Codon engineering for improved antibody expression in mammalian cells. Protein Expr Purif 55, 279-286 crossref(new window)

Beck A, Wurch T, Bailly C and Corvaia N (2010) Strategies and challenges for the next generation of therapeutic antibodies. Nat Rev Immunol 10, 345-352 crossref(new window)

del Val IJ, Kyriakopoulos S, Kontoravdi C et al (2012) Application of Quality by Design paradigm to the manufacture of protein therapeutics; in Glycosylation, Petrescu S (ed.), ISBN: 978-953-51-0771-2, InTech, DOI: 10.5772/ 50261