Sequence and Specificity Analysis of Recombinant Human Fab Anti‐Rh D Isolated by Phage Display

Abstract

These Fab fragments show potential for the development of a new generation of therapeutic anti-Rh D reagents.

A single recombinant anti-RhD IgG prevents RhD immunization: association of RhD-positive red blood cell clearance rate with polymorphisms in the FcγRIIA and FcγIIIA genes

“…In support of this hypothesis, it has been shown in vitro that MonoRho can competitively inhibit the binding of Rhophylac to the RhD antigen on RBCs. 9 Our results confirm other clinical data demonstrating that monoclonal antibodies generated a slower RBC clearance rate than BLOOD, 1 JUNE 2004 ⅐ VOLUME 103, NUMBER 11 For personal use only. on May 13, 2018.…”

“…Its development was based on RhD-specific phage isolated from phage display libraries and subsequent construction of the full-length human IgG1. 2,9 MonoRho recognizes a discontinuous epitope on loops 3, 4, and 6 of the RhD protein (Miescher et al 2 ; and S.M., unpublished data, July 2002). Clinical material was produced in a 200-L batchfermentation process, purified, and supplied in ready-to-use syringes containing 300 g antibody in 1 mL solution.…”

Natural anti-FcεRIα autoantibodies may interfere with diagnostic tests for autoimmune urticaria

“…Reverse transcription of RNA into cDNA and PCR were performed as described [39]. Subsequently, cloning of light chains and heavy chain fragments into the pMVS vector, transformation into XL1-Blue cells, and production of phages were carried out [40,41].…”

Expression and functional analysis of recombinant scFv and diabody fragments with specificity for human RhD

“…Previous studies directed at characterization of anti-D antibodies have revealed a restricted pattern of germline V gene usage (Thompson et al, 1991;Bye et al, 1992;Thorpe et al, 1997Thorpe et al, , 1998Chang and Siegel, 1998), the most commonly used being VH gene segments DP50 and DP63, the JH6 gene, the VlDPL16 and the Jl2/3 genes. In addition, all of the 13 phage-display derived anti-D Fab fragments generated by Miescher and coworkers were based on the DP50 germline gene (Miescher et al, 1998). Our own analysis of rearranged anti-D VH genes contained in GenBank database (accession numbers: AF107231, AF107233, AF107234, AF107236, AF107237, AF107238, AF107239, AF107241, HSA010443, HSA010447, HSU43754, HSU43756, HSU43758, HSU43759, HSU43760, HSU43761, HSU43762; unpublished) demonstrated a bias towards the usage of the DP50 and DP63 (7/17) and JH6 (9/17) germline gene segments.…”