title: Comparison of two functional kappa light-chain transcripts amplified from a hybridoma.
abstract: Three heavy-chain and three kappa (κ)-chain transcripts were amplified from hybridoma cells secreting a monoclonal antibody (mAb) against transferrin receptor. Sequence analysis via IMGT/V-QUEST yielded the functional/aberrant prediction. Two functional κ-chain transcripts, Vκ2 and Vκ3, and one functional VH1 were revealed. Comprehensive bioinformatics analyses including sequence alignment, phylogenetic tree, somatic hypermutation prediction, and three-dimensional-molecular structure modeling were used to predict the origin of the two κ-chain transcripts. The results of bioinformatics analysis suggest that Vκ3 is derived from the myeloma partner of the hybridoma; Vκ2 is derived from B-cell. Functional transcripts VH1 and Vκ2 and Vκ3 were then used to construct two chimeric antibodies chi-C2 (Vκ2-VH1) and chi-C3 (Vκ3-VH1), respectively. Antigen-binding experiments showed that only chi-C2 remained the same affinity as its parental mAb. Possible explanations for the coexistence of two functional κ-chain transcripts and the different affinity of the two chimeric antibodies are discussed.
authors: Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, PR China.
authors: Juan
authors: J
authors: Yang
authors: Huifen
authors: H
authors: Zhu
authors: Zheng
authors: Z
authors: Tan
authors: Fengrong
authors: F
authors: He
authors: Xiaoxu
authors: X
authors: Sun
authors: Yi
authors: Y
authors: Hong
authors: Heyu
authors: H
authors: Hu
authors: Jing
authors: J
authors: Bian
authors: Yu
authors: Y
authors: Lin
authors: Ping
authors: P
authors: Lei
authors: Guanxin
authors: G
authors: Shen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D016466:CHO Cells; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D019295:Computational Biology / Q000379:methods; D003412:Cricetulus; D005803:Genes, Immunoglobulin / Q000235:genetics / Q000276:immunology; D018922:HL-60 Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000235:genetics / Q000276:immunology; D010802:Phylogeny; D011990:Receptors, Transferrin / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment
chemical_list: D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D011990:Receptors, Transferrin