Anti-CD73 humanized antibody
A VH-CDR3, VL-CDR3 technology, applied in the field of biomedicine, can solve the problems of clinical treatment limitations, no CD73-specific targeting antibodies on the market, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0219] Antibody preparation
[0220] Any method suitable for producing monoclonal antibodies can be used to produce CD73 antibodies of the invention. For example, animals can be immunized with linked or naturally occurring CD73 protein or fragments thereof. Suitable immunization methods may be used, including adjuvants, immunostimulants, repeated booster immunizations, one or more routes may be used.
[0221] Any suitable form of CD73 can be used as an immunogen (antigen) for producing non-human antibodies specific to CD73 and screening the biological activity of the antibodies. Immunogens can be used alone or in combination with one or more immunogenicity enhancers known in the art. Immunogens can be purified from natural sources, or produced in genetically modified cells. The DNA encoding the immunogen can be genomic or non-genomic in origin (eg cDNA). DNA encoding the immunogen can be expressed using a suitable genetic vector, including but not limited to adenoviral vec...
Embodiment 1
[0263] Embodiment 1 Preparation of anti-human CD73 monoclonal antibody
[0264] This example mainly describes the sequence of anti-human CD73 single chain antibody obtained by mouse immunization and phage display. Balb / C mice were immunized with recombinant human CD73 protein (C446, Novoprotein). After the first immunization, booster immunization was done every 14 days for a total of 4 immunizations. Mouse serum was collected for antibody titer evaluation. To evaluate the effective mice, take B cells, extract RNA for reverse transcription, and construct a phage display library. Pack the recombinant human CD73 protein at 1-5 μg / ml, add the displayed phages, and perform phage library screening. After the screening, the unwashed phages are collected and infected with host bacteria to obtain the first round of screening library. Follow this method Second and third rounds of screening. After the screening is completed, phage-ELISA identification is carried out, and those identifi...
Embodiment 2
[0269] Example 2 Humanization of CQ137
[0270] Using structural simulation and rational design to analyze the human framework closest to CQ137, a series of humanized antibodies were obtained. The VH and VL of the humanized sequence were respectively constructed on vectors containing hIgG1 and kappa light chain constant regions, transfected into Freestyle 293F cells, the supernatant was collected, and purified on protein A column to obtain the desired antibody protein. The obtained CQ137 humanized antibodies are numbered 137-4, 137-5, 137-6, 137-7, 137-8, 137-9, and their VH and VL sequences are as follows:
[0271] 137-4 VH (SEQ ID NO.: 9)
[0272] QVQLVQSGAEVKKPGSSVKVSCKAS GYSFTGYY IHWVRQAPGQGLEWMGR INPYNGAT TYNQNFKDRVTITVDKSTSTAYMELSSLRSEDTAVYYC ARFHYGAVDY WGQGTLVTVSS
[0273] 137-4 VL (SEQ ID NO.: 15)
[0274] DIQMTQSPSSLSASVGDRVTITCKAS QDINTY LTWYQQKPGKAPKTLIY RAN ILVDGVPSRFSGSGSGTDFTLTISSLQPEDFATYYC LQYEEFPLT FGGGTKVEIKR
[0275] 137-5 VH (SEQ ID NO.: 10)...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More