✦ LIBER ✦

Parallel, high-throughput purification of recombinant antibodies for in vivo cell assays

✍ Scribed by David Bannister; Andrew Wilson; Leah Prowse; Meg Walsh; Rob Holgate; Lutz Jermutus; Trevor Wilkinson

Publisher: John Wiley and Sons
Year: 2006
Tongue: English
Weight: 357 KB
Volume: 94
Category: Article
ISSN: 0006-3592
DOI: 10.1002/bit.20914

No coin nor oath required. For personal study only.

✦ Synopsis

We describe a method for high-throughput, parallel purification of secreted proteins to analyse large numbers of protein samples in cell-based assays for the discovery of protein therapeutics. The procedure is generic and capable of 96 parallel purifications and compatible, in both yield and purity, with a wide assay range. By optimising expression and purification steps as well as using novel hardware, in particular a chromatography press capable to purify target proteins from viscous media, we exemplify the process for the generation of single-chain Fv antibody fragments (scFv) and the purification of full-length IgG. The described process can operate robustly with a throughput of over 2,000 samples per month.

📜 SIMILAR VOLUMES

Biomimetic macroporous hydrogel scaffold

Biomimetic macroporous hydrogel scaffolds in a high-throughput screening format for cell-based assays

✍ Maria B. Dainiak; Irina N. Savina; Isabella Musolino; Ashok Kumar; Bo Mattiasson 📂 Article 📅 2008 🏛 American Institute of Chemical Engineers 🌐 English ⚖ 523 KB

High-Throughput Screening for Methionyl-

High-Throughput Screening for Methionyl-tRNA Synthetases That Enable Residue-Specific Incorporation of Noncanonical Amino Acids into Recombinant Proteins in Bacterial Cells

✍ Tae Hyeon Yoo; David A. Tirrell 📂 Article 📅 2007 🏛 John Wiley and Sons 🌐 English ⚖ 247 KB 👁 2 views

High-Throughput Screening for Methionyl-

High-Throughput Screening for Methionyl-tRNA Synthetases That Enable Residue-Specific Incorporation of Noncanonical Amino Acids into Recombinant Proteins in Bacterial Cells

✍ Tae Hyeon Yoo; David A. Tirrell 📂 Article 📅 2007 🏛 John Wiley and Sons 🌐 English ⚖ 247 KB 👁 2 views

Detection of a decrease in green fluores

Detection of a decrease in green fluorescent protein fluorescence for the monitoring of cell death: An assay amenable to high-throughput screening technologies

✍ Ann-Muriel Steff; Marylène Fortin; Chantal Arguin; Patrice Hugo 📂 Article 📅 2001 🏛 John Wiley and Sons 🌐 English ⚖ 223 KB 👁 3 views

A Reporter Gene Assay for High-Throughpu

A Reporter Gene Assay for High-Throughput Screening of G-Protein-Coupled Receptors Stably or Transiently Expressed in HEK293 EBNA Cells Grown in Suspension Culture

✍ Yves Durocher; Sylvie Perret; Eric Thibaudeau; Marie-Helene Gaumond; Amine Kamen 📂 Article 📅 2000 🏛 Elsevier Science 🌐 English ⚖ 184 KB

We describe in detail a robust, sensitive, and versatile functional assay for G-protein-coupled receptors (GPCRs) expressed in human embryonic kidney (HEK) 293-EBNA (Epstein-Barr virus nuclear antigen) (designated 293E) cells. The ability to grow these cells in suspension, in conjunction with the us

Recombinant antibody toxins specific for

Recombinant antibody toxins specific for ErbB2 and EGF receptor inhibit the in vitro growth of human head and neck cancer cells and cause rapid tumor regression in vivo

✍ Marc Azemar; Mathias Schmidt; Friederike Arlt; Pierre Kennel; Burkhard Brandt; A 📂 Article 📅 2000 🏛 John Wiley and Sons 🌐 French ⚖ 130 KB 👁 2 views

## Overexpression of the ErbB2 and epidermal growth factor receptor (EGFR) tyrosine kinases is frequently observed in squamous cell carcinomas of the head and neck, and has been correlated with shorter overall survival. By immunoblot analysis, we have found EGFR and ErbB2 expression in 6 out of 6 e