| 摘要 |
High-affinity monoclonal antibodies, wherein the affinity is characterised b y: (i) incubating first and second samples of the antibody in antigen-coated microtitre plate wells at a concentration chosen to be within the linear par t of a standard curve at pH 7.2 for 1 hour at 37 ~C; (ii) removing unbound antibody from both samples; (iii) incubating the first sample with PBS at pH 7.2 for 1 hour at 37 ~C, and reducing the pH of the second sample to pH 3 or below and incubating for 1 hour at 37 ~C; (iv) removing unbound antibody fro m both samples; (v) incubating both samples with anti-antibody alkaline phosphatase-conjugate for 1 hour at 37 ~C; (vi) removing unbound conjugate from both samples; and (vii) adding PNPP substrate to the samples, measuring the absorbance of the samples at 405 nm, and determining the amount of antibody bound to antigen, wherein the amount bound in the second sample is > 50 % of that of the first sample.
|
