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Lambda fabselect

Manufactured by GE Healthcare
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The Lambda FabSelect is a lab equipment product designed for protein purification. It is a chromatography system that utilizes protein A or protein G affinity media to selectively capture and purify antibodies and antibody fragments from complex samples.

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Lab products found in correlation

Pcdna3, by Thermo Fisher Scientific (2 mentions) Hitrap kappaselect, by GE Healthcare (1 mentions)

4 protocols using lambda fabselect

1

Isolation and Purification of Human mAbs

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Human mAbs were isolated previously from healthy adults who were naturally infected with a GII.4 Sydney virus in 2013 using an established human B cell hybridoma technique (27 (link)). Here, cloned hybridoma cell lines that had been cryopreserved were thawed and placed into culture and expanded gradually from 48-well plates to 12-well plates, T-25, T-75, and eventually to multiple T-225 flasks for each cell line. Following 4 weeks of incubation at 37°C, supernatant from the T-225 flasks was harvested and filtered through a 0.4-µm filter. The supernatant was purified using column chromatography, specifically HiTrap KappaSelect and Lambda FabSelect affinity resins (GE Healthcare Life Sciences).
Ford-Siltz L.A., Tohma K., Alvarado G.S., Kendra J.A., Pilewski K.A., Crowe JE J.r, & Parra G.I. (2022). Cross-reactive neutralizing human monoclonal antibodies mapping to variable antigenic sites on the norovirus major capsid protein. Frontiers in Immunology, 13, 1040836.
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2

Production and Purification of Antibodies

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For the production of purified full-length IgG1 antibodies, the VHand VL genes were cloned into the pCDNA3.1+ (Invitrogen)mammalian expression vector containing either the human IgG1 constant region gene, the human kappa light chain constant region gene or the lambda light chain constant region gene using standard recombinant DNA technology30 (link). For the production of the purified nCoV396Fab antibody, a stop codon TGA was introduced after the sequence (5′-TCTTGTGACAAA-3′), which encodes the amino acid residues SCDK, just before the hinge of the human IgG1 constant region40 (link). Recombinant IgG1 antibodies and the nCoV396 antibody Fab fragments were produced in 293F cells cultured in serum-free medium by co-transfection with the generated IgG1 full-length or Fab heavy- and light chain gene expression plasmid pairs using polyethylenimine41 (link). Full-length IgG1 antibodies were purified by using Protein A column chromatography as described previously30 (link). The nCoV396 antibody Fab used for the crystal structure was purified by Lambda FabSelect, an affinity resin designed for the purification of human Fab with a lambda light chain (GE Healthcare)40 (link).
Kang S., Yang M., He S., Wang Y., Chen X., Chen Y.Q., Hong Z., Liu J., Jiang G., Chen Q., Zhou Z., Zhou Z., Huang Z., Huang X., He H., Zheng W., Liao H.X., Xiao F., Shan H, & Chen S. (2021). A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation. Nature Communications, 12, 2697.
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3

Crystallization of Fab-IL-2 Complex

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Antibody F5111 Fab was expressed by transient transfection in HEK293 cells and purified by LambdaFabSelect chromatography (GE Life Sciences). F5111 Fab was mixed with a 1.5-fold excess of IL-2, and the complex was purified by fast protein liquid chromatography on a Superdex 200 column. The purified Fab−IL-2 complex was concentrated to 10 mg ml−1 as measured at 280 nm using an extinction coefficient of 1.49 ml mg−1 cm−1. The complex was mixed with an equal volume of precipitant solution (100 mM sodium citrate, pH 5.0; 20% polyethylene glycol 6000) and crystallized by sitting-drop vapor diffusion over a reservoir of precipitant solution. Prismatic crystals formed within one week. Crystals were cryoprotected by addition of glycerol to 30% (v/v), collected and rapidly cooled by plunging into liquid nitrogen.
Trotta E., Bessette P.H., Silveria S.L., Ely L.K., Jude K.M., Le D.T., Holst C.R., Coyle A., Potempa M., Lanier L.L., Garcia K.C., Crellin N.K., Rondon I.J, & Bluestone J.A. (2018). A human anti-IL-2 antibody that potentiates regulatory T cells by a structure-based mechanism. Nature medicine, 24(7), 1005-1014.
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4

Production and Purification of IgG1 Antibodies

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For production of purified full-length IgG1 antibodies, the VHDJH and VL genes were cloned into the pCDNA3.1+ (Invitrogen) mammalian expression vector containing either the human IgG1 constant region gene, the human lambda light-chain constant region, or the lambda light-chain constant region gene using standard recombinant DNA technology (45 (link)). For the production of purified nCoV617Fab antibody, a stop codon TGA was introduced after the sequence (5′-TCTTGTGACAAA-3′) encoding amino acid residues, SCDK, just before the hinge of the human IgG1 constant region (46 (link)). Recombinant IgG1 antibodies and the nCoV617Fab antibody were produced in 293F cells cultured in serum-free medium by cotransfection with the generated full-length IgG1 or Fab heavy- and light-chain gene expression plasmid pairs using polyethylenimine (47 (link)). Full-length IgG1 antibodies were purified by using protein A column chromatography as described previously (45 (link)). The nCoV617Fab antibody used for the crystal structure was purified by Lambda FabSelect, an affinity resin designed for the purification of human Fab with a lambda light chain (GE Healthcare) (46 (link)).
Yang M., Li J., Huang Z., Li H., Wang Y., Wang X., Kang S., Huang X., Wu C., Liu T., Jia Z., Liang J., Yuan X., He S., Chen X., Zhou Z., Chen Q., Liu S., Li J., Zheng H., Liu X., Li K., Yao X., Lang B., Liu L., Liao H.X, & Chen S. (2021). Structural Basis of a Human Neutralizing Antibody Specific to the SARS-CoV-2 Spike Protein Receptor-Binding Domain. Microbiology Spectrum, 9(2), e01352-21.
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