Example 4

The antibody affinity was analyzed using BIACORE Biomacromolecular Interaction Device (GE). The anti-human IgG-Fab antibody (Abcam) was conjugated on a chip, the anti-human IgG-Fab antibody was used to capture the SG001 antibody or the BMS01 positive antibody, the antibody concentration was set to 1 μg/mL, and the sample injection time was 60-150 seconds; the antigen hPD1-FC was used as a mobile phase, and 6 concentration gradients (3.125, 6.25, 12.5, 25, 50, 100 nM) were used, with a binding time of 120 seconds; the dissociation time was 1200S; 10 mM glycine-hydrochloric acid buffer (pH 2.1) was used for regeneration, for a period of 60S. The results showed that the affinities of the SG001 antibody and the BMS01 positive antibody were 2.15 nM and 1.4 nM, respectively, which were substantially same. The results were shown in Table 1.

TABLE 1
Determination results for the affinities of the SG001
antibody and the BMS01 positive antibody
AssociationDissociationAffinity
constantconstantconstant
AntibodyKa (1/Ms)Kd (1/s)KD (M)
BMS015.594 × 1057.900 × 10−51.412 × 10−10
SG0013.184 × 1056.850 × 10−52.151 × 10−10

US10774144B2. Human programmed cell death 1 receptor antibody, method of preparing same, and use thereof (2020-09-15). HANGZHOU SUMGEN BIOTECHNOLOGY Co., Ltd. [CN]. Inventors: Fan Yang [CN].
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