Primary sequence determination and molecular modelling of the variable region of an antiMUC1 mucin monoclonal antibody.
Polymerase chain reaction (PCR) products representative of the DNA sequence coding for the variable heavy (VH) and the variable light (VL) chains of an antiMUC1 mucin monoclonal antibody, C595, have been produced. These products were cloned, sequenced, and the primary amino acid sequences of the VH and VL regions deduced. The hypervariable complementarity determining regions (CDRs) and framework regions in the heavy and light chains were located, and homologies with canonical forms for the CDR loops L1, L2, L3, H1 and H2 were identified by database searching. The structure for the H3 loop was calculated directly. Computational molecular modelling was accomplished using the fully automated AbM package (Oxford Molecular, Oxford, U.K.). Energy minimisation was performed using the program InsightII (Biosym, San Diego, California, U.S.A.). The investigation provides a basis for the molecular analysis of the antigen binding site of the C595 antibody with the aim to identify key residues and interactions involved in the immune recognition of the C595 antibody defined epitope, which is expressed in the majority of breast and ovarian carcinomas.
Denton
G
G
Cancer Research Laboratory, University of Nottingham, University Park, U.K.
Davies
G M
GM
Scanlon
M J
MJ
Tendler
S J
SJ
Price
M R
MR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics; D001483:Base Sequence; D001666:Binding Sites, Antibody; D004247:DNA / Q000737:chemistry; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D009077:Mucins / Q000276:immunology*; D016133:Polymerase Chain Reaction
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D009077:Mucins; D004247:DNA