Next Article in Journal
Properties of Fluorescent Far-Red Anti-TNF Nanobodies
Previous Article in Journal
ADME Considerations and Bioanalytical Strategies for Pharmacokinetic Assessments of Antibody-Drug Conjugates
Article Menu

Export Article

Open AccessArticle
Antibodies 2018, 7(4), 42; https://doi.org/10.3390/antib7040042

Enhancers Improve the AID-Induced Hypermutation in Episomal Vector for Antibody Affinity Maturation in Mammalian Cell Display

1,2,†
,
1,2,†
,
1,2
,
3
,
4
,
5,* and 1,2,*
1
Key Laboratory for Protein and Peptide Pharmaceuticals, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
2
University of Chinese Academy of Sciences, No.19(A) Yuquan Road, Shijingshan District, Beijing 100049, China
3
Hefei Institutes of Physical Science, Chinese Academy of Sciences, Institute of Technical Biology and Agriculture Engineering, 350 Shushanhu Road, Hefei 230031, Anhui, China
4
National Institute of Metrology, No.18, Bei San Huan Dong Lu, Chaoyang Dist, Beijing 100101, China
5
Department of Civil and Environmental Engineering, Northeastern University, Boston, MA 02115, USA
These authors contributed equally to the work.
*
Authors to whom correspondence should be addressed.
Received: 18 November 2018 / Revised: 10 December 2018 / Accepted: 11 December 2018 / Published: 13 December 2018
Full-Text   |   PDF [2644 KB, uploaded 18 December 2018]   |  

Abstract

The induction of somatic hypermutation (SHM) in various cell lines by activation-induced cytidine deaminase (AID) has been used in protein-directed selection, especially in antibody affinity maturation. Several antibody affinity maturation systems based on mammalian cells have been developed in recent years, i.e., 293T, H1299, Raji and CHO cells. However, the efficiency of in vitro AID-induced hypermutation is low, restricting the application of such systems. In this study, we examined the role of Ig and Ek enhancers in enhancing SHM in the episomal vector pCEP4 that expresses an anti-high mobility group box 1 (HMGB1) full-length antibody. The plasmid containing the two enhancers exhibited two-fold improvement of mutation rate over pCEP4 in an AID expression H1299 cell line (H1299-AID). With the engineered episomal vector, we improved the affinity of this antibody in H1299-AID cells by 20-fold. View Full-Text
Keywords: activation-induced cytidine deaminase (AID); somatic hypermutation (SHM); episomal vector; enhancer activation-induced cytidine deaminase (AID); somatic hypermutation (SHM); episomal vector; enhancer
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Chen, C.; Wang, J.; Zhao, Y.; Chen, S.; Hu, Z.; Chen, L.; Hang, H. Enhancers Improve the AID-Induced Hypermutation in Episomal Vector for Antibody Affinity Maturation in Mammalian Cell Display. Antibodies 2018, 7, 42.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Antibodies EISSN 2073-4468 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top