humanization of antibodies service丨Zoonbio

Summary

Human anti-mouse antibody response (HAMA response) can be produced when non-human antibodies enter human body, which will affect the safety and therapeutic effect of antibodies in clinical application. Humanization of antibody is to convert most or all of the sequence of antibody into human sequence through gene engineering operation and recombinant expression. On the basis of ensuring affinity and specificity of antibody, the heterology of antibody to human is reduced to the greatest extent.

Using CDR transplantation technology and computer-aided structural simulation design, Zoonbio can humanize mouse-derived monoclonal antibody, rabbit-derived monoclonal antibody and alpaca nano-antibody to ensure that the degree of humanization of antibody is more than 90%, and the affinity is higher than or equal to chimeric antibody.


monoclonal antibody customization platform	monoclonal antibody sequencing platform	humanized antibody design platform	recombinant antibody expression platform

·preparation of monoclonal antibodies of mice and rabbits ·construction of phage display library ·preparation of alpaca nanoantibodies	·hybridoma cells based on PCR amplification antibody sequencing ·complete sequencing of antibody proteins based on high resolution mass spectrometry	·three-dimensional structure modeling of antibody based on database ·selection of humanized antibody template based on homology ·identification of amino acid residues that maintain the conformation of CDR ·analysis of key amino acid recovery mutation	·mature mammalian cell expression system ·20L-1000L high throughput expression equipment

antibody sequencing	cycle	delivery of data and materials
monoclonal antibody sequencing	2 weeks	antibody sequence
humanized design
● three-dimensional structure modeling of antibodies ● classical residue analysis ● human template search and humanized sequence generation ● recovery catastrophe analysis based on classical residue analysis ● combination of humanized antibody VH and VL (20 pairs)	1-2 weeks	report on humanized design of antibodies
vector construction
● codon optimization ● gene synthesis ● cloning heavy and light chain genes of antibodies to expression vectors, respectively ● endotoxin-free plasmid extraction	1-2 weeks	gene synthesis report commercial expression vectors containing antibody heavy chain and light chain respectively escherichia coli DH10 containing commercially expressed plasmids
small-scale expression of transient transfection
● transient transfection of HEK293 cells ● expression test, condition optimization ● antibody purification ● antibody analysis and identification (SDS-PAGE) and UV analysis	2-3 weeks	the purified humanized antibodies with the best affinity of 1-3 strains were 0.2-0.5 mg per strain antibody expression report
ELISA and Octet/Biacore determination of affinity	1-2 weeks	affinity determination report
Remarks:1.Customers need to provide antibody sequences and antigen proteins 2.For high purity or low endotoxin antibodies, please contact customer service for special instructions.

Summary

Humanized Antibody Technology Platform

service flow