Comparison of the biological properties of two anti-mucin-1 antibodies prepared for imaging and therapy.
A comparison was made between the murine anti-MUC1 antibody BC2 (which reacts with the peptide epitope APDTR) and the "humanised" antibody hCTMO1 from CellTech, which reacts with the MUC1 epitope RPAP. Preliminary studies demonstrated that hCTMO1 was a "good" antibody whereas BC2 was not. Various parameters were determined and conclusions reached. (a) Affinity: the affinity of hCTMO1 was 2.60 x 10(7) M(-1) and that of BC2 was 1.36 x 10(7) M(-1); we did not consider these numbers to be substantially different, although hCTMO1 was clearly of higher affinity than BC2. (b) On/off rate at 4 degrees C: both antibodies bound effectively to the MUC-1 transfectant MOR5-CF2; the association rate for hCTMO1 was 3.8 times that of BC2 and the dissociation rate for BC2 was twice as fast as that of hCTMO1. (c) On/off rates at 37 degrees C: at 37 degrees C the association rate for hCTMO1 was greater than that of BC2. (d) Internalization: hCTMO1 was also more efficient at internalising bound antibody; 70% of bound hCTMO1 was internalised, whilst 6% of bound BC2 was internalised. From these studies it was clear that, while hCTMO1 was of similar affinity to BC2, the faster uptake and internalisation and lower off rate indicated that it was likely to be a superior antibody; this was proven in vivo. (e) Localisation: hCTMO1 bound much better in vivo than BC2 (68% compared to 28%). (f) Therapeutic experiments: BC2-idarubicin conjugates were essentially ineffective in eradicating tumours in mice whereas hCTMO1-idarubicin had a dramatic effect on breast cancer tumour cells growing in mice. We conclude that the simple measurements on/off rates and internalisation at 37 degrees C are the most important parameters to use to determine antibody effectiveness, prior to embarking on clinical studies.
Pietersz
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GA
Austin Research Institute, Heidelberg, Victoria, Australia.
Wenjun
L
L
Krauer
K
K
Baker
T
T
Wreschner
D
D
McKenzie
I F
IF
D016475:3T3 Cells; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism* / Q000494:pharmacology*; D000941:Antigens / Q000378:metabolism; D000939:Epitopes / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D007457:Iodine Radioisotopes; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D018396:Mucin-1 / Q000276:immunology*; D013696:Temperature; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D007457:Iodine Radioisotopes; D018396:Mucin-1