The Versatility of Framework Regions of Chicken VH and VL to Mutations
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To identify the interchangeability of VH and VL framework region (FR) residues, we artificially introduced random mutations at all residue positions in a chicken monoclonal antibody, which has only one functional VH and Vλ gene. When we classified the amino acids into 5 groups by their physicochemical properties, all FR residues could be replaced by another group except L23 (C), H36 (W), H86 (D), H104 (G), and H106 (G). Eighty-two (50.9%), 48 (29.8%), 17 (10.6%), and 9 FR residues (5.6%) could be replaced by 4, 3, 2, and 1 group(s), individually, without significant loss of reactivity. We also confirmed a similar level of versatility with 2 different chicken antibodies. This high level of versatility on FR residues has not been predicted because it has not been observed in the 150 chicken antibodies that we previously generated or in the 1,269 naïve chicken VH sequences publically available. In conclusion, chicken antibody FR residues are highly interchangeable and this property can be applied for improving the physicochemical property of antibody including thermal stability, solubility and viscosity.
Abstract
To identify the interchangeability of VH and VL framework region (FR) residues, we artificially introduced random mutations at all residue positions in a chicken monoclonal antibody, which has only one functional VH and Vλ gene. When we classified the amino acids into 5 groups by their physicochemical properties, all FR residues could be replaced by another group except L23 (C), H36 (W), H86 (D), H104 (G), and H106 (G). Eighty-two (50.9%), 48 (29.8%), 17 (10.6%), and 9 FR residues (5.6%) could be replaced by 4, 3, 2, and 1 group(s), individually, without significant loss of reactivity. We also confirmed a similar level of versatility with 2 different chicken antibodies. This high level of versatility on FR residues has not been predicted because it has not been observed in the 150 chicken antibodies that we previously generated or in the 1,269 naïve chicken VH sequences publically available. In conclusion, chicken antibody FR residues are highly interchangeable and this property can be applied for improving the physicochemical property of antibody including thermal stability, solubility and viscosity.
참고문헌 (18)
- Cohen RM, "Somatic hypermutation targeting is influenced by location within the immunoglobulin V region", Mol Immunol, 48, pp.1477~1483 (2011)[DOI]
- Balint RF, "Antibody engineering by parsimonious mutagenesis", Gene, 137, pp.109~118 (1993)[DOI]
- Barbas CF 3rd, "Selection and evolution of high-affinity human anti-viral antibodies", Trends Biotechnol, 14, pp.230~234 (1996)[DOI]
- Haidar JN, "A universal combinatorial design of antibody framework to graft distinct CDR sequences: a bioinformatics approach", Proteins, 80, pp.896~912 (2012)[DOI]
- Wu L, "Fundamental characteristics of the immunoglobulin VH repertoire of chickens in comparison with those of humans, mice, and camelids", J Immunol, 188, pp.322~333 (2012)[DOI]
- Barbas CF, "Phage Display: a Laboratory Manual" (2001), Cold Spring Harbor Laboratory Press
- Kabat EA, "Sequences of Proteins of Immunological Interest", U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, ~ (1991)
- Kim HY, "Production and characterization of chimeric monoclonal antibodies against Burkholderia pseudomallei and B. mallei using the DHFR expression system", PLoS One, 6, pp.e19867~ (2011)[DOI]
- Tsurushita N, "Humanization of a chicken anti-IL-12 monoclonal antibody", J Immunol Methods, 295, pp.9~19 (2004)[DOI]
- Murray F, "Stereotyped patterns of somatic hypermutation in subsets of patients with chronic lymphocytic leukemia: implications for the role of antigen selection in leukemogenesis", Blood, 111, pp.1524~1533 (2008)
- Wiens GD, "Somatic mutation in VH complementarity-determining region 2 and framework region 2: differential effects on antigen binding and Ig secretion", J Immunol, 159, pp.1293~1302 (1997)
- Wagner SD, "Codon bias targets mutation", Nature, 376, pp.732~ (1995)[DOI]
- Shapiro GS, "Predicting regional mutability in antibody V genes based solely on di- and trinucleotide sequence composition", J Immunol, 163, pp.259~268 (1999)
- Yang J, "Substitution of surface-exposed framework residues alters secretion of recombinant fusion protein Fv", tumor necrosis factor in Escherichia coli, 1999, pp.48, 327~332
- Rodriguez-Rodriguez ER, "A single mutation in framework 2 of the heavy variable domain improves the properties of a diabody and a related single-chain antibody", J Mol Biol, 423, pp.337~350 (2012)[DOI]
- Tanha J, "Improving solubility and refolding efficiency of human V(H)s by a novel mutational approach", Protein Eng Des Sel, 19, pp.503~509 (2006)[DOI]
- Tangri S, "Rationally engineered proteins or antibodies with absent or reduced immunogenicity", Curr Med Chem, 9, pp.2191~2199 (2002)[DOI]
- Shih HH, "An ultra-specific avian antibody to phosphorylated tau protein reveals a unique mechanism for phosphoepitope recognition", J Biol Chem, 287, pp.44425~44434 (2012)[DOI]