Fab Expression and Purification in E. coli

Fabs were expressed and purified as reported previously (57 (link)). Briefly, genes encoding antibody clones were cloned into a vector that expresses a Fab with the Avi-tag at the C-terminus of the heavy chain. Fabs were produced in E. coli strain 55244 (ATCC) and purified using a HiTrap Protein G affinity column (Cytiva). Purity >90% was confirmed using SDS-PAGE.

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Hattori T., Maso L., Araki K.Y., Koide A., Hayman J., Akkapeddi P., Bang I., Neel B.G, & Koide S. (2022). Creating MHC-Restricted Neoantigens with Covalent Inhibitors That Can Be Targeted by Immune Therapy. Cancer Discovery, 13(1), 132-145.

Publication 2022

HiTrap Protein G affinity column

Antibody E coli Genes Protein g Sds page Strain Vector

Corresponding Organization : New York University

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Variable analysis

independent variables

Genes encoding antibody clones

dependent variables

Purity of Fabs (confirmed using SDS-PAGE)

control variables

Avi-tag at the C-terminus of the heavy chain
E. coli strain 55244 (ATCC) for Fab production
HiTrap Protein G affinity column (Cytiva) for Fab purification

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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