AN100226m

Antibody Name: AN100226m (VH SGIIC | VL kV)
References: Leger, O.J.P., Yednock, T.A., Tanner, L., Horner, H.C., Hines, D.K., Keen, S., Saldanha, J., Jones, S.T., Fritz, L.C. and Bendig, M.M. "Humanization of a mouse antibody against human alpha-4 integrin: A potential therapeutic for the treatment of multiple sclerosis" Hum. Antibod. (1997) 8:3-16.
Donor Antibody: mouse
Acceptor Antibody: Human IgG1 | k (later Celltech produced IgG4)
Antigen: human α4 integrin on very late antigen-4 (VLA-4, α4B1, CD49d/CD29) adhesion molecule expressed on the surface of lukocytes which binds to vascular cell adhesgion molecule-1 (VCAM-1) which is expressed on cytokine-stimulated endothelial cells.
Laboratory: Athena Neurosciences, CA; MRC Collaborative Centre, UK
Design: Best-fit strategy plus a model of V regions to analyse possible mutations.
Frameworks: VH 21/28'CL | VL REI (with modifications L104V Q105E T107K)
CDRs: All 6 VH and VL, Kabat Definition.
Backmutations: VH YTFT27-30FNIK R71A (positions 27 and 29 not important)
VL K45R Y49H V58 T69R
Binding: "Almost identical affinity" for human Jurkat T cells (with IgG4 isotype). Inhibits CNS inflammation in Experimental Autoimmune Encephalomyelitis (EAE) in animal studies.
Expression: Transient expression in coscells. Stable expressing cell line made at Celltech, UK.
Comment: Tested backmutations at 38:H 40:H 44:H and found they were not important, even if they were all positively charged. Loss of potential glycosylation site at 74:L with no effect on binding.
Clinical Indication: Acute and chronic inflammatory conditions (eg multiple sclerosis, rheumatoid arthritis, asthma, psoriasis and allergy).

explanation of symbols and abbreviations

José Saldanha © 1997-8. Birkbeck College, London WC1E 7HX.