Characterisation and internalisation of recombinant humanised HMFG-1 antibodies against MUC1

The humanised HMFG-1 immunoglobulin has been extensively developed as a clinical immunotherapeutic agent for MUC1 expressing tumours. We have constructed a single-chain Fv (scFv) and Fab fragment from this antibody and shown that both these species retain their specificity for MUC1. The scFv was les...

Full description

Saved in:
Bibliographic Details
Published in:British journal of cancer 2005-11, Vol.93 (11), p.1257-1266
Main Authors: Pericleous, L M, Richards, J, Epenetos, A A, Courtenay-Luck, N, Deonarain, M P
Format: Article
Language:English
Subjects:
Amino acids
Antibodies
Antibodies, Monoclonal - immunology
Antigens
Binding Sites, Antibody
Breast Neoplasms - pathology
Cancer research
Glycoproteins
Humans
Immunoglobulin Fab Fragments - immunology
Immunoglobulins
Kinases
Kinetics
Ligands
Medical research
Mucin-1 - immunology
Proteins
Translational Therapeutics
Tumor Cells, Cultured
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The humanised HMFG-1 immunoglobulin has been extensively developed as a clinical immunotherapeutic agent for MUC1 expressing tumours. We have constructed a single-chain Fv (scFv) and Fab fragment from this antibody and shown that both these species retain their specificity for MUC1. The scFv was less stable and less soluble than the Fab. Detailed analyses of the binding kinetics of the whole IgG and Fab fragment show that the affinity for MUC1 synthetic peptides is low (approximately 100 nM for the IgG and 10 muM for the Fab), with particularly low but similar dissociation rate constants (0.031-0.095 s(-1)). Binding to native antigen on the cell surface is over two orders of magnitude better. Confocal immunofluorescence microscopy shows that both the IgG and Fab are internalised rapidly (the IgG is internalised within 15 min) and colocalise to early endosomes. This work provides an appreciation of the binding, internalising and trafficking kinetics, important for the development of future therapeutics based on this antibody.
ISSN:0007-0920
1532-1827
DOI:10.1038/sj.bjc.6602847