Monoclonal Antibody Production from Hybridoma Cells

The mAb-hERG1 (MCK Therapeutics; Florence, Italy; patent number IT1367861) was produced by secretion from an hybridoma cell line (named A7) obtained from mice cell fusion and able to secrete the monoclonal antibody (Guasti et al., 2008 (link); Lastraioli et al., 2020 (link); Lottini et al., 2021 (link)). Hybridoma cells are routinely maintained in DMEM (Euroclone) supplemented with Glutamine 10 mM and 5% Fetal Clone serum (Euroclone). Supernatant is harvested and purified by affinity chromatography using AKTA Pure (Cytiva) mounted with Protein A columns (Cytiva).

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Santini L., Duranti C., Palandri C., Giammarino L., Musumeci M., Carlucci L., Capitani C., Colasurdo R., Recchia F., Cerbai E., Coppini R, & Arcangeli A. (2023). Cardiac safety assessment of a novel recombinant bispecific antibody targeting the ether-à-go-go related gene 1 (hERG1)-β1 integrin macromolecular complex. Frontiers in Pharmacology, 14, 1237431.

Publication 2023

DMEM Glutamine AKTA Pure Protein A columns

Affinity chromatography Cell fusion Cell line Cells Clone Fetal Glutamine Hybridoma Mice Monoclonal antibody Protein a Secretion Serum Therapeutics

Corresponding Organization : University of Florence

Other organizations : Scuola Superiore Sant'Anna, University of Siena

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Variable analysis

independent variables

Hybridoma cell line (named A7) obtained from mice cell fusion and able to secrete the monoclonal antibody

dependent variables

Production and secretion of the monoclonal antibody (mAb-hERG1)

control variables

Hybridoma cells are routinely maintained in DMEM (Euroclone) supplemented with Glutamine 10 mM and 5% Fetal Clone serum (Euroclone)

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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