Affinity-based chromatography is a powerful and specific separation method used for bioactive compound isolation and purification employing the binding affinity between immune complexes. Specific monoclonal antibodies or patient immunoglobulin have been used to capture human or mouse tumor tissue-derived antigens, HLA-restricted tumor-specific antigens, and neoantigens. Following the capture process, the proteins bound to chromatography are eluted and evaluated by various other methods. Creative Biolabs has extensive antibody development and production experience and offers the chromatography assay for antibody-dependent cancer epitope screening and identification.
Fig.1 Immunoaffinity chromatography for protein purification.1
With highly efficient chromatography technology, Creative Biolabs provides various types of chromatography assays for antigen and antibody purification and cancer epitope analysis with a particular emphasis on immunoaffinity chromatography and size exclusion chromatography (SEC).
✔ Antibody Development and Purification
Fig.2 Different formats of antibodies.2
✔ Antigen Protein Purification
Antigen protein could be purified from two pathways, immunoaffinity chromatography, and receptor-ligand-based affinity chromatography assay.
High-specificity monoclonal antibodies are used to purify antigens based on binding affinity.
Fig.3 Workflow of immunoaffinity chromatography for antigen purification. (Creative Biolabs)
A variety of methods are supplied to immobilize and couple antibodies to the matrix with the right direction and site.
The effective elution conditions ensure antigen peptides elute from the column and keep the column easy for regeneration.
Fig.4 The principal and working steps of immunoaffinity chromatography for antigen purification.3
Biotinylated monoclonal antibodies are incubated with the antigen protein mixture, and the formed antigen-antibody complex is purified by an avidin column. Finally, the bound antigens are separated using a proper elution buffer.
Fig.5 Specific antigen-antibody complex purification using chromatography.4
✔ Antigen-antibody Complex Purification
SEC, also known as gel filtration chromatography, is widely used to collect highly purified, homogeneous antigen-antibody complex from a mixture based on molecule size.
✔ Techniques and Assays for Epitope Peptide Analysis
The eluted antigen binders can be used to immunize animals to acquire antisera and then examined by multiple assays.
Amino Acid Sequencing: Liquid chromatography and mass spectrometry assay.
Purity Tests: Immunoprecipitation assay and SDS-PAGE.
Specificity Tests: Western blot, ELISA, and antibody mediated cross blocking assay.
3D Structure study: X-ray crystallization, small-angle scattering, and nuclear magnetic resonance (NMR) assay.
Please contact us and consult the service of your interest.
For Research Use Only | Not For Clinical Use
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