Optimized dual assay for the transgenes selection and screening in CHO cell line development for recombinant protein production

Objective To develop a simple robust methodology of screening multiple CHO cell clones secreting recombinant proteins to assess their specific productivity. Results We developed a dual assay based on immunoassay measurements of a recombinant protein expression combined with staining of viable cells...

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Bibliographic Details
Published in:Biotechnology letters 2019-09, Vol.41 (8-9), p.929-939
Main Authors: Beketova, Elena V., Ibneeva, Liliia R., Abdulina, Yulia A., Dergousova, Elena A., Filatov, Vladimir L., Kozlovsky, Stanislav V., Shilov, Evgeny S., Datskevich, Petr N., Rozov, Fedor N.
Format: Article
Language:English
Subjects:
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biotechnology
Cloning
Growth rate
Heterogeneity
Immunoassay
Life Sciences
Microbiology
Original Research Paper
Productivity
Protein expression
Proteins
Screening
Transfection
Transgenes
Workflow
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Summary:Objective To develop a simple robust methodology of screening multiple CHO cell clones secreting recombinant proteins to assess their specific productivity. Results We developed a dual assay based on immunoassay measurements of a recombinant protein expression combined with staining of viable cells with resazurin. Following this approach, colonies can be simultaneously assessed for cell growth rate and for production of a recombinant protein. Combination of these two assays enables to estimate productivity of a recombinant protein per cell from the very early stages of a cell line development process (CLD) and exclude poor producers from further steps. Comparison of the dual assay with a standard CLD protocol followed by only analysis of protein expression level showed at least 10–20% increase in the amount of clones that can be included into pool of high-producers at early stages. This shortens duration of a typical CLD scheme from 23 to 19 weeks. Conclusions Our method: (i) allows to include into workflow clones that demonstrate slow growth during single cell cloning but producing high amounts of a target protein, which otherwise would be lost in standard protocols of cells screening; (ii) can be applied for testing of DNA vectors for transfection and protein production; (iii) can be used for monitoring the heterogeneity of cell population and analysis of stable pools productivity.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-019-02711-4