Comparison between hybridoma and Fab/phage anti-RhD: Their V gene usage and pairings

Our 11 anti-RhD's in conjunction with 37 previously published RhD antibodie s, produced by hybridoma technology were analysed for germline gene usage a nd restriction in V-H and V-L pairings. The 17 V-H germline genes used by t he hybridoma anti-RhD IgG were derived from 4 VH families (VH1, VH2, VH3 an d VH4). Eighteen kappa chains were restricted to only 5 germline genes from only 2 V kappa families (V kappa1 and V kappa3). However, the 13 lambda ch ains were not as restricted, using 10 Vh germline genes from 4 families(VXI , V lambda2, V lambda3 and V lambda8). Fifty six unique Fab/phage anti-RhD were also analysed. In all cases the Fab/phage VH germline genes were deriv ed from the VH3 family (41/41). The 29 kappa chains were restricted to 3 ge rmline genes primarily from V kappa1 (97%) and the 13 lambda chains used 10 V lambda germline genes from 5 families (V lambda1, V lambda2, V lambda3, V lambda4 and V lambda7). The V-H germline genes of the Fab/phage were rest ricted 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- kappa germline genes used by the IgG (DPK9). However, the repertoire of the V-lambda germline genes used in these two systems is different. with analy sis showing greater diversity in V-lambda gene usage with 8 unique germline genes used by 76% Fab/phage compared to 4 unique genes used by 46% of the IgG hybriboma anti-RhD.