Method for detecting coupling distribution of intermediate T-MCC of antibody coupling medicine T-DM1
A T-DM1, antibody coupling technology, applied in the field of medicine, can solve problems such as interference, affecting accuracy, background fluorescence, etc., and achieve the effects of easy operation, cost saving, and product quality assurance
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0055] Example 1 Carry out reduced CE-SDS analysis to T-MCC synthesized by different production processes and corresponding T-DM1
[0056] Reduction of the test solution preparation:
[0057] T-MCC sample 1 (18mg / mL): Take 50 μL of T-MCC sample 1, add 175 μL of water, and dilute to 4 mg / mL. According to the preparation method of the reduced test solution, take 25 μL of diluted T-MCC sample 1, add 70 μL of SDS sample buffer, 5 μL of β-mercaptoethanol, and mix well. Heat at 65°C for 10min.
[0058] T-MCC sample 2 (19.5 mg / mL): Take 50 μL of T-MCC sample 2, add 194 μL of water, and dilute to 4 mg / mL. Take 25 μL, add 70 μL of sample buffer, 5 μL of β-mercaptoethanol, and mix well. Heat at 65°C for 10min.
[0059] T-MCC sample 3 (17.6mg / mL): Take 50 μL of T-MCC sample 3, add 170 μL of water, and dilute to 4 mg / mL. Take 25 μL, add 70 μL of sample buffer, 5 μL of β-mercaptoethanol, and mix well. Heat at 65°C for 10min.
[0060] T-DM1 sample 1 (22.0 mg / mL): Take 50 μL of T-DM1 ...
Embodiment 2
[0081] Reduction of the test solution preparation:
[0082] Test solution 1: Take 50 μL of T-MCC sample 1, add 175 μL of water, and dilute to 4 mg / mL. Take 25 μL of diluted T-MCC sample 1, add 70 μL of SDS sample buffer, 5 μL of β-mercaptoethanol, and mix well. Heat at 65°C for 10min.
[0083] Test solution 2: Take 50 μL of T-MCC sample 1, add 175 μL of water, and dilute to 4 mg / mL. Take 25 μL of diluted T-MCC sample 1, add 70 μL of SDS sample buffer, 5 μL of dithiothreitol, and mix well. Heat at 65°C for 10min.
[0084] Test solution 3: Take 50 μL of T-MCC sample 1, add 175 μL of water, and dilute to 4 mg / mL. Take 25 μL of diluted T-MCC sample 1, add 70 μL of SDS sample buffer, 5 μL of β-mercaptoethanol, and mix well. Heat at 60°C for 10min.
[0085] Test solution 4: take 50 μL of T-MCC sample 1, add 175 μL of water, and dilute to 4 mg / mL. Take 25 μL of diluted T-MCC sample 1, add 70 μL of SDS sample buffer, 5 μL of β-mercaptoethanol, and mix well. Heat at 70°C for 10...
Embodiment 3
[0094] Reduction of the test solution preparation:
[0095] Test solution: Take 50 μL of T-MCC sample 1, add 175 μL of water, and dilute to 4 mg / mL. Take 25 μL of diluted T-MCC sample 1, add 70 μL of SDS sample buffer, 5 μL of β-mercaptoethanol, and mix well. Heat at 65°C for 10min.
[0096] Capillary cartridge installation: Use 50 μm ID uncoated fused silica capillary (such as uncoated capillary 50 μm ID from SCIEX), cut to a total length of 30 cm and an effective length of 20 cm. The detection window is equipped with hole plug No. 2 (100 μm × 200 μm).
[0097] Assay 1: PA800plus is analyzed using capillary electrophoresis. The capillary temperature is 20°C, the sample chamber temperature is 10°C, and the detection wavelength is 220nm. Rinse the capillary with 0.1M NaOH solution at 70psi pressure for 3min, wash the capillary with 0.1M HCl solution at 70psi pressure for 1min, wash the capillary with ultrapure water at 70psi pressure for 1min, and fill it with gel buffer at...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More