Construction of a reshaped HMFG1 antibody and comparison of its fine specificity with that of the parent mouse antibody.
A human antibody with milk mucin specificity was obtained by transferring the complementarity determining regions (CDR) of the mouse antibody HMFG1 onto carefully selected human framework regions. The resulting reshaped human antibody, HuHMFG1, showed no difference in relative affinity for its antigen compared with the parent mouse HMFG1. The minimum epitope recognized by both the mouse and reshaped antibodies was demonstrated by epitope mapping to be identical, and consists of the tetramer PDTR. In a replacement net analysis, in which each of the amino acids was replaced in turn with the 19 other residues, it was determined that mouse HMFG1 and HuHMFG1 reacted with this series of synthetic peptides in an equivalent manner, indicating retention of identical fine specificity in the HuHMFG1 antibody. In contrast to other published reports, this was achieved without involvement of any framework residues in the binding site transfer. These data demonstrate that if well-matching human framework regions are employed grafting the CDR only can be sufficient to confer desired specificities to human antibodies and can, indeed, provide human analogues of mouse antibodies with virtually indistinguishable affinities and fine specificities relative to the mouse parent antibodies.
Verhoeyen
M E
ME
Unilever Research, Colworth Laboratory, Bedford, U.K.
Saunders
J A
JA
Price
M R
MR
Marugg
J D
JD
Briggs
S
S
Broderick
E L
EL
Eida
S J
SJ
Mooren
A T
AT
Badley
R A
RA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D000939:Epitopes / Q000032:analysis; D006801:Humans; D051379:Mice; D008895:Milk, Human / Q000276:immunology*; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D004247:DNA