High‐throughput and automation advances for accelerating single‐cell cloning, monoclonality and early phase clone screening steps in mammalian cell line development for biologics production

Tejwani, Vijay; Chaudhari, Minal; Rai, Toyaj; Sharfstein, Susan T.

doi:10.1002/btpr.3208

Cited by 11 publications

(7 citation statements)

References 64 publications

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“…However, the low single-cell encapsulation efficiencies and physical damage to cells have seriously hindered its wide application. 35 In this study, CellCelector nanowell array, which combines a high-content imaging cytometry system and high-precision single-cell-picking robotics, 36 was used for hapten-specific rabbit ASC selection. The roofless nanowell structure permits rapid and direct cell retrieval by capillary and the flexible modification endows the monitoring of secreted antibodies based on a tailor-made and effective assay.…”

Section: Resultsmentioning

confidence: 99%

A rare monoclonal antibody discovery based on indirect competitive screening of a single hapten-specific rabbit antibody secreting cell

et al. 2022

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Section: Resultsmentioning

confidence: 99%

A rare monoclonal antibody discovery based on indirect competitive screening of a single hapten-specific rabbit antibody secreting cell

et al. 2022

Analyst

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show abstract

“…However, the low single-cell encapsulation efficiencies and physical damage to cells have seriously hindered its wide application 28 . In this study, CellCelector nanowell array, which combines high-content imaging cytometry system and high-precision single-cell-picking robotics 29 , for hapten-specific rabbit ASC selection. The roofless nanowell structure permits rapid and direct cell retrieval by capillary and the flexible modification endows the monitoring of secreted antibody based on a tailor-made and effective assay.…”

Section: Resultsmentioning

confidence: 99%

“…Droplet-based microfluidics capsulizing cells into aqueous droplets holds great potential for functional screening of ASCs at the single-cell level. However, the low single-cell encapsulation efficiencies and physical damage to cells have seriously hindered its wide application cytometry system and high-precision single-cell-picking robotics 29 , for hapten-specific rabbit ASC selection. The roofless nanowell structure permits rapid and direct cell retrieval by capillary and the flexible modification endows the monitoring of secreted antibody based on a tailor-made and effective assay.…”

Section: Selection Of Single Cap-specific Asc By the Csmnmentioning

confidence: 99%

Rare Monoclonal Antibody Discovery Based on Indirect Competitive Screening of Single Hapten-specific Rabbit Antibody Secreting Cell

Li¹,

Li²,

et al. 2022

Preprint

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Rare antibody that is able to tolerate physio-chemical factors is preferred and highly demanded in diagnosis and therapy. Rabbit monoclonal antibodies (RmAbs) are distinguished owing to their high affinity and stability. However, the efficiency and availability of traditional methods for RmAb discovery are limited, especially for small molecules. Here, we present an indirect competitive screening method in nanowells, named CSMN, for single rabbit antibody secreting cells (ASCs) selection with 20.6 h and proposed an efficient platform for RmAb production against small molecule with 5.8 days for the first time. Chloramphenicol (CAP) as an antibacterial agent has the great threats for public health. We applied the CSMN to select CAP-specific ASCs and produced one high affinity RmAb, surprisingly showing extremely halophilic properties with an IC50 of 0.08 ng mL-1 in saturated salt solution which has as yet not been shown by other antibodies. Molecular dynamic simulation showed that the negatively charged surface improved the stability of the RmAb structure with additional disulfide bonds compared with mouse antibody. Moreover, the reduced solvent accessible surface area of the binding pocket increased the interactions of RmAb with CAP in a saturated salt solution. Furthermore, the RmAb was used to develop an immunoassay for the detection of CAP in real biological samples with simple pretreatment, shorter assay time, and higher sensitivity. The results demonstrated that the practical and efficient CSMN is suitable for rare RmAb discovery against small molecules.

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“…Novel cell line screening methods mostly rely on high-throughput technology to improve the screening efficiency. Moreover, several high throughput methods for CLD are listed, and Tejwani et al (2021) give more details in the development of these technologies.…”

Section: Other Screening Methodsmentioning

confidence: 99%

Screening Strategies for High-Yield Chinese Hamster Ovary Cell Clones

Yang

Zhang

Xiao

et al. 2022

Front. Bioeng. Biotechnol.

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Chinese hamster ovary (CHO) cells are by far the most commonly used mammalian expression system for recombinant expression of therapeutic proteins in the pharmaceutical industry. The development of high-yield stable cell lines requires processes of transfection, selection, screening and adaptation, among which the screening process requires tremendous time and determines the level of forming highly productive monoclonal cell lines. Therefore, how to achieve productive cell lines is a major question prior to industrial manufacturing. Cell line development (CLD) is one of the most critical steps in the production of recombinant therapeutic proteins. Generation of high-yield cell clones is mainly based on the time-consuming, laborious process of selection and screening. With the increase in recombinant therapeutic proteins expressed by CHO cells, CLD has become a major bottleneck in obtaining cell lines for manufacturing. The basic principles for CLD include preliminary screening for high-yield cell pool, single-cell isolation and improvement of productivity, clonality and stability. With the development of modern analysis and testing technologies, various screening methods have been used for CLD to enhance the selection efficiency of high-yield clonal cells. This review provides a comprehensive overview on preliminary screening methods for high-yield cell pool based on drug selective pressure. Moreover, we focus on high throughput methods for isolating high-yield cell clones and increasing the productivity and stability, as well as new screening strategies used for the biopharmaceutical industry.

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High‐throughput and automation advances for accelerating single‐cell cloning, monoclonality and early phase clone screening steps in mammalian cell line development for biologics production

Cited by 11 publications

References 64 publications

A rare monoclonal antibody discovery based on indirect competitive screening of a single hapten-specific rabbit antibody secreting cell

A rare monoclonal antibody discovery based on indirect competitive screening of a single hapten-specific rabbit antibody secreting cell

Rare Monoclonal Antibody Discovery Based on Indirect Competitive Screening of Single Hapten-specific Rabbit Antibody Secreting Cell

Screening Strategies for High-Yield Chinese Hamster Ovary Cell Clones

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