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Papain resin

Manufactured by Thermo Fisher Scientific
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Papain resin is a chromatographic medium used for the purification and separation of proteins and other biomolecules. It is composed of papain, a proteolytic enzyme, immobilized on a solid support matrix. The primary function of papain resin is to facilitate the capture and purification of target proteins from complex mixtures through affinity-based separation techniques.

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Lab products found in correlation

Fab preparation kit, by Thermo Fisher Scientific (2 mentions) Tris glycine sds page, by Thermo Fisher Scientific (2 mentions) Pepsin, by Thermo Fisher Scientific (1 mentions)

3 protocols using papain resin

1

Fab Fragment Production and Purification

Cited 1 time
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Fab fragments were expressed with a His-tag heavy chain expression vector and co-transfected with a light chain vector in Expi293F cells. Fab fragments were also produced by papain digestion with a Fab preparation kit (ThermoFisher, Cat. 44985) according to manufacturer’s protocol. In brief, 0.5-1.0 mg of IgG1 antibodies were mixed with 125 μL papain resin (ThermoFisher, Cat. 20341) for 5 h in the digestion buffer provided, containing 20 mM cysteine, pH 7.4. Undigested antibody and Fc fragments were removed by incubating digested products with a protein A column overnight at 4°C, then collecting the Fab-containing flow-through. Fabs were analyzed by 4%–20% Tris-Glycine SDS-PAGE (ThermoFisher, Cat. XP04200BOX). C12C9 and C12A2 Fabs were prepared by expressing Fabs with a 3C-cleavable histag that was removed using 3C protease (Pierce) following Talon resin purification, as described (Schmidt et al., 2015 (link)).
Tong P., Gautam A., Windsor I.W., Travers M., Chen Y., Garcia N., Whiteman N.B., McKay L.G., Storm N., Malsick L.E., Honko A.N., Lelis F.J., Habibi S., Jenni S., Cai Y., Rennick L.J., Duprex W.P., McCarthy K.R., Lavine C.L., Zuo T., Lin J., Zuiani A., Feldman J., MacDonald E.A., Hauser B.M., Griffths A., Seaman M.S., Schmidt A.G., Chen B., Neuberg D., Bajic G., Harrison S.C, & Wesemann D.R. (2021). Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike. Cell, 184(19), 4969-4980.e15.
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2

Fab Fragment Production and Purification

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Fab fragments were expressed with a His-tag heavy chain expression vector and co-transfected with a light chain vector in Expi293F cells. Fab fragments were also produced by papain digestion with a Fab preparation kit (ThermoFisher, Cat. 44985) according to manufacturer’s protocol. In brief, 0.5–1.0 mg of IgG1 antibodies were mixed with 125 μl papain resin (ThermoFisher, Cat. 20341) for 5 hours in the digestion buffer provided, containing 20 mM cysteine, pH 7.4. Undigested antibody and Fc fragments were removed by incubating digested products with a protein A column overnight at 4 °C, then collecting the Fab-containing flow-through. Fabs were analyzed by 4–20 % Tris-Glycine SDS-PAGE (ThermoFisher, Cat. XP04200BOX). C12C9 and C12A2 Fabs were prepared by expressing Fabs with a 3C-cleavable histag that was removed using 3C protease (Pierce) following Talon resin purification, as described (Schmidt et al., 2015 (link)).
Tong P., Gautam A., Windsor I.W., Travers M., Chen Y., Garcia N., Whiteman N.B., McKay L.G., Storm N., Malsick L.E., Honko A.N., Lelis F.J., Habibi S., Jenni S., Cai Y., Rennick L.J., Duprex W.P., McCarthy K.R., Lavine C.L., Zuo T., Lin J., Zuiani A., Feldman J., MacDonald E.A., Hauser B.M., Griffths A., Seaman M.S., Schmidt A.G., Chen B., Neuberg D., Bajic G., Harrison S.C, & Wesemann D.R. (2021). Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike. Cell, 184(19), 4969-4980.e15.
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3

Monoclonal Antibody Fragment Generation

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F(ab’)2 and Fab fragments were generated by digestion of monoclonal antibodies on immobilized pepsin and papain resin (both Thermo Fisher Scientific), respectively, followed by subsequent purification on a protein A column to remove Fc fragments and undigested antibody.
Seitz C.M., Mittelstaet J., Atar D., Hau J., Reiter S., Illi C., Kieble V., Engert F., Drees B., Bender G., Krahl A.C., Knopf P., Schroeder S., Paulsen N., Rokhvarguer A., Scheuermann S., Rapp E., Mast A.S., Rabsteyn A., Schleicher S., Grote S., Schilbach K., Kneilling M., Pichler B., Lock D., Kotter B., Dapa S., Miltenyi S., Kaiser A., Lang P., Handgretinger R, & Schlegel P. (2021). Novel adapter CAR-T cell technology for precisely controllable multiplex cancer targeting. Oncoimmunology, 10(1), 2003532.
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