Cloning of a hamster anti-mouse CD79B antibody sequences and identification of a new hamster immunoglobulin lambda constant IGLC gene region

Abstract

Anti-CD79 antibodies have been effective at targeting B cell lymphoma cells and depleting B cells in animal models. In order to engineer recombinant antibodies with additional effector functions in mice, we cloned and sequenced the full-length cDNAs of the heavy and light chain of a hamster anti-mouse CD79B antibody. Although hamster antibodies represent a unique source of monoclonal antibodies against mouse, rat, and human antigens, sequence information of hamster immunoglobulins (IG) is sparse. Here, we report a new hamster (Cricetulus migratorius) IG lambda constant (IGLC) gene region that is most homologous to mouse IGLC2 and IGLC3.

Fig. 1

Nucleotide sequence and deduced translation of HM79-16 heavy chain. The predicted leader region cleavage site is indicated by “//”. The polyadenylation signal is underlined. Conserved amino acid cysteine 23 (1st-CYS) and cysteine 104 (2nd-CYS) are highlighted (Lefranc, 2011b; Lefranc et al., 2003; Lefranc et al., 2005). The full-length sequences have been deposited in GenBank (accession number KC294425).

Fig. 2

Nucleotide sequence and deduced translation of HM79-16 light chain. The predicted leader region cleavage site is indicated by “//”. The polyadenylation signal is underlined. Conserved amino acid cysteine 23 (1st-CYS) and cysteine 104 (2nd-CYS) are highlighted (Lefranc, 2011b; Lefranc et al., 2003; Lefranc et al., 2005). The full-length sequences have been deposited in GenBank (accession number KC294424).

Fig. 3

IMGT protein displays for V domains of HM79-16 and HL4E10. VH and V-LAMDA sequences of HM79-16 (KC294425 and KC294424) and HL4E10 (HM369133 and HM369134) are aligned using IMGT/DomainGapAlign (Ehrenmann and Lefranc, 2011).

Fig. 4

IMGT protein displays for C regions of HM79-16 and HL4E10. The heavy gamma C region comprises three domains (CH1, CH2, CH3), the hinge between CH1 and CH2, and the CHS (present at the C terminal end of a secreted chain). CH1, hinge, CH2, CH3, CHS of HM79-16 and HL4E10 are presented successively (top). The light lambda C region comprises a single C domain (C-LAMBDA). C-LAMBDA of HM79-16 (KC294424), HL4E10 (HM369134), and mouse IGLC1 (J00587), IGLC2 (J00595), IGLC3 (J00585) are shown (bottom). Sequences are aligned using IMGT/DomainGapAlign (Ehrenmann and Lefranc, 2011).

Fig. 5

IMGT Collier de Perles of the C-LAMDA for HM79-16 (A) and HL4E10 (B). IMGT Colliers de Perles were obtained using the IMGT/Collier-de-Perles tool (Ehrenmann et al., 2011), accessed from IMGT/DomainGapAlign.

Fig. 6

Phylogenetic tree of the IGLC regions of different species. The tree was generated by the neighbor-joining method using ClustalW2 program (Larkin et al., 2007). The bar (0.1) indicates the genetic distance. The IGLC regions of various species were obtained from IMGT^®, the international ImMunoGeneTics information system^® (http://www.imgt.org), IMGT/GENE-DB (Giudicelli et al., 2005; Lefranc et al., 2009). The name and accession numbers (in parentheses) are: human IGLC1 (J00252), IGLC2 (X06875), IGLC3 (J00254), IGLC6 (J03011), IGLC7 (X51755); mouse IGLC1 (J00587), IGLC2 (J00595), IGLC3 (J00585); rabbit IGLC1 (M12388), IGLC2 (M12761), IGLC3 (M12762), IGLC4 (M12763), IGLC5 (M23227), IGLC6 (M23229); Rat IGLC1 to 4 (AC111360). The standardized IMGT 6-letter abbreviations are used: Homsap (Homo sapiens, human), Musmus (Mus musculus, house mouse), Orycun (Oryctolagus cuniculus, rabbit), Ratnor (Rattus norvegicus, Norway rat), Crimig (Cricetulus migratorius, Armenian hamster).