Comparison between Hybridoma and Fab/phage Anti-RhD: Their V Gene Usage and Pairings

Abstract

Our 11 anti-RhD's in conjunction with 37 previously published RhD antibodies, produced by hybridoma technology were analysed for germline gene usage and restriction in V_H and V_L pairings. The 17 V_H germline genes used by the hybridoma anti-RhD IgG were derived from 4 VH families (VH1, VH2, VH3 and VH4). Eighteen kappa chains were restricted to only 5 germline genes from only 2 Vκ families (Vκ1 and κ3). However, the 13 lambda chains were not as restricted, using 10 V_λ germline genes from 4 families (Vλ1, Vλ2, Vλ3 and Vλ8). Fifty six unique Fab/phage anti-RhD were also analysed. In all cases the Fab/phage V_H germline genes were derived from the VH3 family (41/41). The 29 kappa chains were restricted to 4 germline genes primarily from Vκ1 (97%) germline genes from 5 families (Vλ1, Vλ2, Vλ3, Vλ4 and Vλ7). The V_H germline genes of the Fab/phage were restricted to 4 of the 17 used by the hybridoma anti-RhD IgG (DP46, DP49, DP50 and DP77). Ninety percent of the Fab/phage were restricted to 1 of the 5 V_κ germline genes used by the IgG (DPK9). However, the repertoire of the V_λ germline genes used in these two systems is different, with analysis showing greater diversity in V_λ gene usage with 8 unique germline genes used by 76% Fab/phage compared to 4 unique genes used by 46% hybriboma anti-RhD.