Imperial College London

ProfessorRobinShattock

Faculty of MedicineDepartment of Infectious Disease

Chair in Mucosal Infection and Immunity
 
 
 
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Contact

 

+44 (0)20 7594 5206r.shattock

 
 
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Location

 

453Wright Fleming WingSt Mary's Campus

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Summary

 

Publications

Citation

BibTex format

@article{Aw:2018:10.1016/j.pep.2018.03.013,
author = {Aw, R and McKay, P and Shattock, R and Polizzi, K},
doi = {10.1016/j.pep.2018.03.013},
journal = {Protein Expression and Purification},
pages = {43--50},
title = {A systematic analysis of the expression of the anti-HIV VRC01 antibody in Pichia pastoris through signal peptide optimization},
url = {http://dx.doi.org/10.1016/j.pep.2018.03.013},
volume = {149},
year = {2018}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - Pichia pastoris (Komagataella phaffi) has been used for recombinant protein production for over 30 years with over 5000 proteins reported to date. However, yields of antibody are generally low. We have evaluated the effect of secretion signal peptides on the production of a broadly neutralizing antibody (VRC01) to increase yield. Eleven different signal peptides, including the murine IgG1 signal peptide, were combinatorially evaluated for their effect on antibody titer. Strains using different combinations of signal peptides were identified that secreted approximately 2-7 fold higher levels of VRC01 than the previous best secretor, with the highest yield of 6.50 mg L-1 in shake flask expression. Interestingly it was determined that the highest yields were achieved when the murine IgG1 signal peptide was fused to the light chain, with several different signal peptides leading to high yield when fused to the heavy chain. Finally, we have evaluated the effect of using a 2A signal peptide to create a bicistronic vector in the attempt to reduce burden and increase transformation efficiency, but found it to give reduced yields compared to using two independent vectors.
AU - Aw,R
AU - McKay,P
AU - Shattock,R
AU - Polizzi,K
DO - 10.1016/j.pep.2018.03.013
EP - 50
PY - 2018///
SN - 1046-5928
SP - 43
TI - A systematic analysis of the expression of the anti-HIV VRC01 antibody in Pichia pastoris through signal peptide optimization
T2 - Protein Expression and Purification
UR - http://dx.doi.org/10.1016/j.pep.2018.03.013
UR - http://hdl.handle.net/10044/1/58612
VL - 149
ER -