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A procedure for producing an anti-AXL nanobody in E. coli.
- Source :
-
Protein Expression & Purification . Jul2023, Vol. 207, pN.PAG-N.PAG. 1p. - Publication Year :
- 2023
-
Abstract
- As one of the receptors of the TAM family, AXL plays a vital role in stem cell maintenance, angiogenesis, immune escape of viruses and drug resistance against tumors. In this study, the truncated extracellular segment containing two immunoglobulin-like domains of human AXL (AXL-IG), which has been confirmed to bind growth arrest specific 6 (GAS6) by structural studies [1], was expressed in a prokaryotic expression system and then purified. Immunizing camelid with the purified AXL-IG as antigen could lead to the production of unique nanobodies composed of only variable domain of heavy chain of heavy-chain antibody (VHH), which are around 15 kD and stable. We screened out a nanobody A-LY01 specific binding to AXL-IG. We further determined the affinity of A-LY01 to AXL-IG and revealed that A-LY01 could specifically recognize full-length AXL on the surface of HEK 293T/17 cells. Our study provides appropriate support for the development of diagnostic reagents and antibody therapeutics targeting AXL. • AXL-IG protein was expressed in E. coli WK6. • A nanobody phage library was constructed. • A nanobody A-LY01 specifically binds to AXL was screened out by phage display technology. [ABSTRACT FROM AUTHOR]
- Subjects :
- *ESCHERICHIA coli
*STEM cells
*IMMUNOGLOBULINS
*DRUG resistance
*BACTERIOPHAGES
Subjects
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 207
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 163422563
- Full Text :
- https://doi.org/10.1016/j.pep.2023.106268