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Rapid high throughput cdna synthesis platform

Manufactured by Twist Bioscience
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The Rapid high-throughput cDNA synthesis platform is a lab equipment designed for the rapid and high-throughput generation of complementary DNA (cDNA) from RNA samples. The core function of this platform is to efficiently convert RNA into cDNA, which is a crucial step in various molecular biology and genomics applications.

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Lab products found in correlation

50 ml mini bioreactor tubes, by Corning (2 mentions) Amicon ultra 4 50 kda centrifugal filter units, by Merck Group (2 mentions) Hitrap mabselect sure, by Cytiva (1 mentions)

5 protocols using rapid high throughput cdna synthesis platform

1

High-Throughput Production of Monoclonal Antibodies

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Sequences of selected mAbs were synthesized using a rapid high-throughput cDNA synthesis platform (Twist Bioscience) and subsequently cloned into an IgG1 monocistronic expression vector (designated as pTwist) for mammalian cell culture mAb secretion. This vector contains an enhanced 2A sequence and GSG linker that allows simultaneous expression of mAb heavy and light chain genes from a single construct upon transfection 25 (link).
Gilchuk P., Bombardi R.G., Erasmus J.H., Tan Q., Nargi R., Soto C., Abbink P., Suscovich T.J., Durnell L.A., Khandhar A., Archer J., Liang J., Fouch M.E., Davidson E., Doranz B.J., Jones T., Larson E., Ertel S., Granger B., Fuerte-Stone J., Roy V., Broge T., Linnekin T.C., Linde C.H., Gorman M.J., Nkolola J., Alter G., Reed S.G., Barouch D.H., Diamond M.S., Crowe JE J.r., Van Hoeven N., Thackray L, & Carnahan R. (2020). Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics. Nature biomedical engineering, 4(11), 1030-1043.
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2

High-Throughput Mammalian mAb Production

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The most somatically mutated representatives from each clonal family and members of all public clonotypes were synthesized using a rapid high-throughput cDNA synthesis platform (Twist Bioscience) and subsequently cloned into an IgG1 monocistronic expression vector (designated as pTwist-mCis_G1) for mAb secretion from mammalian cell culture. This vector contains an enhanced 2A sequence and GSG linker that allows simultaneous expression of mAb heavy- and light-chain genes from a single construct upon transfection.63 (link) We performed transfections of ExpiCHO cell cultures using the Gibco ExpiCHO Expression System and protocol for 50 mL mini bioreactor tubes (Corning) as described by the vendor. Culture supernatants were purified using HiTrap MabSelect SuRe (Cytiva) on a 24-column parallel protein chromatography system (Protein Biosolutions). Purified mAbs were buffer-exchanged into PBS, concentrated using Amicon Ultra-4 50-kDa centrifugal filter units (Millipore Sigma) and stored at 4°C until use.
(1996). Proceedings of the National Academy of Sciences of the United States of America, 93(25), 14993.
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3

Rapid High-Throughput mAb Production

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Sequences of selected mAbs were synthesized using a rapid high-throughput cDNA synthesis platform (Twist Bioscience) and subsequently cloned into an IgG1 monocistronic expression vector (designated as pTwist-mCis_G1) for mammalian cell culture mAb secretion. This vector contains an enhanced 2A sequence and GSG linker that allows simultaneous expression of mAb heavy and light chain genes from a single construct upon transfection47 .
Zost S.J., Gilchuk P., Chen R.E., Case J.B., Reidy J.X., Trivette A., Nargi R.S., Sutton R.E., Suryadevara N., Chen E.C., Binshtein E., Shrihari S., Ostrowski M., Chu H.Y., Didier J.E., MacRenaris K.W., Jones T., Day S., Myers L., Lee F.E., Nguyen D.C., Sanz I., Martinez D.R., Rothlauf P.W., Bloyet L.M., Whelan S.P., Baric R.S., Thackray L.B., Diamond M.S., Carnahan R.H, & Crowe JE J.r. (2020). Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein. Nature medicine, 26(9), 1422-1427.
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4

High-Throughput cDNA Synthesis and Monoclonal Antibody Production

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Sequences of mAbs were synthesized using a rapid high-throughput cDNA synthesis platform (Twist Bioscience) and subsequently cloned into an IgG1 monocistronic expression vector (designated as pTwist-mCis_G1) for mAb secretion from mammalian cell culture. This vector contains an enhanced 2A sequence and GSG linker that allows simultaneous expression of mAb heavy- and light-chain genes from a single construct upon transfection (Chng et al., 2015 (link)). We performed transfections of ExpiCHO cell cultures using the GIBCO ExpiCHO Expression System and protocol for 50mL mini bioreactor tubes (Corning) as described by the vendor. Culture supernatants were purified using HiTrap MabSelect SuRe (Cytiva, formerly GE Healthcare Life Sciences) on a 24-column parallel protein chromatography system (Protein Biosolutions). Purified monoclonal antibodies were buffer exchanged into PBS, concentrated using Amicon Ultra-4 50-kDa centrifugal filter units (Millipore Sigma) and stored at 4°c until use.
Chen E.C., Gilchuk P., Zost S.J., Suryadevara N., Winkler E.S., Cabel C.R., Binshtein E., Chen R.E., Sutton R.E., Rodriguez J., Day S., Myers L., Trivette A., Williams J.K., Davidson E., Li S., Doranz B.J., Campos S.K., Carnahan R.H., Thorne C.A., Diamond M.S, & Crowe JE J.r. (2021). Convergent antibody responses to the SARS-CoV-2 spike protein in convalescent and vaccinated individuals. Cell Reports, 36(8), 109604.
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5

High-Throughput Production of Monoclonal Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Sequences of selected mAbs were synthesized using a rapid high-throughput cDNA synthesis platform (Twist Bioscience) and subsequently cloned into an IgG1 monocistronic expression vector (designated as pTwist) for mammalian cell culture mAb secretion. This vector contains an enhanced 2A sequence and GSG linker that allows simultaneous expression of mAb heavy and light chain genes from a single construct upon transfection 25 (link).
Gilchuk P., Bombardi R.G., Erasmus J.H., Tan Q., Nargi R., Soto C., Abbink P., Suscovich T.J., Durnell L.A., Khandhar A., Archer J., Liang J., Fouch M.E., Davidson E., Doranz B.J., Jones T., Larson E., Ertel S., Granger B., Fuerte-Stone J., Roy V., Broge T., Linnekin T.C., Linde C.H., Gorman M.J., Nkolola J., Alter G., Reed S.G., Barouch D.H., Diamond M.S., Crowe JE J.r., Van Hoeven N., Thackray L, & Carnahan R. (2020). Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics. Nature biomedical engineering, 4(11), 1030-1043.
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