PMID- 34920133
OWN - NLM
STAT- MEDLINE
DCOM- 20220217
LR  - 20220217
IS  - 1096-0279 (Electronic)
IS  - 1046-5928 (Linking)
VI  - 192
DP  - 2022 Apr
TI  - Capture and purification of an untagged nanobody by mixed weak cation 
      chromatography and cation exchange chromatography.
PG  - 106030
LID - S1046-5928(21)00213-8 [pii]
LID - 10.1016/j.pep.2021.106030 [doi]
AB  - Nanobodies (Nbs) are single-domain antibodies, which have potential application 
      value in tumor-targeted therapy, immunotherapy, diagnostic probe, and molecular 
      imaging. Typically, Nbs are captured by affinity chromatography via the addition 
      of specific fusion tags at their N or C terminus. Nerve growth factor (NGF), 
      which regulates the growth and development of peripheral and central neurons, 
      maintains neuronal survival and plays a key role in both arthritis and acute and 
      chronic pain. In this study, a method for capture and purification of an untagged 
      Nb (anti-NGF Nb) by mixed weak cation chromatography and cation exchange 
      chromatography was established. First, pH 4.0-5.0 was demonstrated to be the 
      optimal loading condition for Capto MMC to capture anti-NGF by the design of 
      experiments (DOE). Furthermore, high purity and yield products can be obtained at 
      laboratory scale and commercial production scale by adjusting the protein pH. 
      Additionally, direct capture of anti-NGF Nb using Capto MMC without adjusting 
      anti-NGF Nb harvest pH was investigated. The anti-NGF Nb captured by Capto MMC 
      was 67.2% yield, 94.5% monomer purity, and host cell protein (HCP) was reduced 
      from 74,931 ppm to 484 ppm. The anti-NGF Nb that were further purified using 
      Capto S ImpAct achieved 84.5% yield and 99.2% purity and 77 ppm of HCP. The 
      scaling-up process was consistent with the results of the optimized process, 
      further demonstrating the feasibility of this method. This outcome provides a 
      highly promising and competitive alternative to affinity chromatography-based 
      processing strategies for Nbs.
CI  - Copyright (c) 2021 Elsevier Inc. All rights reserved.
FAU - Li, Xin
AU  - Li X
AD  - Key Laboratory of Molecular Pharmacology and Drug Evaluation, Ministry of 
      Education, School of Pharmacy, Yantai University, Yantai, 264005, PR China.
FAU - Liu, Wanhui
AU  - Liu W
AD  - Key Laboratory of Molecular Pharmacology and Drug Evaluation, Ministry of 
      Education, School of Pharmacy, Yantai University, Yantai, 264005, PR China.
FAU - Li, Heshui
AU  - Li H
AD  - Shandong Boan Biotechnology Company Limited, PR China.
FAU - Wang, Xin
AU  - Wang X
AD  - Shandong Boan Biotechnology Company Limited, PR China.
FAU - Zhao, Yanyan
AU  - Zhao Y
AD  - Key Laboratory of Molecular Pharmacology and Drug Evaluation, Ministry of 
      Education, School of Pharmacy, Yantai University, Yantai, 264005, PR China. 
      Electronic address: zhaoyy@luye.com.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20211214
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (Cation Exchange Resins)
RN  - 0 (Single-Chain Antibodies)
SB  - IM
MH  - Adsorption
MH  - Cation Exchange Resins/chemistry
MH  - Chromatography, Ion Exchange/*methods
MH  - Single-Chain Antibodies/chemistry/genetics/*isolation & purification/metabolism
OTO - NOTNLM
OT  - Capto MMC
OT  - Capto S ImpAct
OT  - Capture
OT  - Mixed weak cation chromatography
OT  - Nanobodies
OT  - Purification
EDAT- 2021/12/18 06:00
MHDA- 2022/02/19 06:00
CRDT- 2021/12/17 20:14
PHST- 2021/10/21 00:00 [received]
PHST- 2021/12/09 00:00 [revised]
PHST- 2021/12/10 00:00 [accepted]
PHST- 2021/12/18 06:00 [pubmed]
PHST- 2022/02/19 06:00 [medline]
PHST- 2021/12/17 20:14 [entrez]
AID - S1046-5928(21)00213-8 [pii]
AID - 10.1016/j.pep.2021.106030 [doi]
PST - ppublish
SO  - Protein Expr Purif. 2022 Apr;192:106030. doi: 10.1016/j.pep.2021.106030. Epub 
      2021 Dec 14.