Serological Analysis by Recombinant cDNA Expression Cloning

The serological analysis of recombinant cDNA expression clones originated from the mid-1990s and is a technique for binding a cDNA expression library with a serum phenotype to molecular cloning. By detecting the antibody response in the blood of tumor patients, and screening and searching for tumor antigen genes from tumor cell cDNA expression libraries. Creative Biolabs can provide accurate data analysis and comprehensive technical support about serological analysis by recombinant cDNA expression cloning.

Introduction of SEREX Technology

Over the past 10 years, various types of tumor antigens have been screened using the serological analysis by recombinant cDNA expression cloning (SEREX) technique, including melanoma, kidney cancer, colon cancer, and the like. In recent years, the application of this technology in non-lung neoplastic diseases has also been reported, such as autoimmune diseases, systemic lupus erythematosus, rheumatoid arthritis, and Behcet's disease.

The SEREX technology combines immunological and molecular cloning methods in which phage using cDNA is used to transfect host bacteria. The colonies were then blotted with an NC membrane, and positive clones were screened by sera and hybridization. For antigen screening, the library titer is first determined. If the inoculation is too dense, the phage grows slowly, the plaque is small, and the protein expression is small, and the connected spots have an effect on each other's color development. If the amount of inoculation is too small, and the number of clones will too small, which will reduce the incidence of positive clones. Therefore, the plaque overlap should be selected, the plaque growth should be uniform, and the amount of cDNA library screened must reach 10⁶ clones or more to reflect the structure of the native gene as completely as possible.

Protocol of SEREX Preparation of host bacteria: determination of titer of empty phage and human embryonic stem cell cDNA library. Pretreatment of different serum: membrane absorption method, modified membrane absorption method, direct pretreatment method. Optimization of the closed method. Observe the color depth, background treatment, and false positives Highlights Efficient, short detection cycle, low cost High quality, strong medical background, and rich practical experience High evaluation, excellent technical support

Creative Biolabs provides one-stop customized services, especially in the aspect of SEREX technology. If you want to know more information, please contact us and we will happy to help you.

References

1. Luna‐Coronell, J.A.; et al. The prostate cancer immunome: In silico functional analysis of antigenic proteins from microarray profiling with IgG. Proteomics. 2016, 16(8): 1204-1214.
2. Coronell, J.A.L.; et al. The immunome of colon cancer: functional in silico analysis of antigenic proteins deduced from IgG microarray profiling. Genomics, Proteomics & Bioinformatics. 2018, 16(1): 73-84.

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