Affinity maturation of glucose transporter SLC2A1 antibody 6F12#33 by phage display
Files
Date
2023
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Tartu Ülikool
Abstract
Antibodies against glucose transporter SLC2A1 have high potential is cancer therapy. Thus, this
master's thesis aimed to enhance the affinity of antibody 6F12#33 against glucose transporter
SLC2A1, which can then be used to inhibit the glucose uptake in various cancers, essentially
starving and killing the cancer cells. The lead antibody 6F12#33 was discovered at Icosagen, which
was then used in affinity maturation. Antibody maturation libraries were designed and generated,
leading to the screening of approximately 600 colonies using ELISA. From these 600 clones, eight
promising clones were selected based on signal intensity and sequence similarity. Antibody
sequences were cloned into human IgG1 format, produced in CHO cells, and purified.
Comprehensive assays, including cell proliferation and various binding assays, were conducted to
evaluate their efficacy. Three out of the eight affinity-matured antibodies improved binding affinity
to SLC2A1 compared to the parental which translated to a significant enhancement of their
inhibitory effects. The remaining five antibodies exhibited similar or decreased inhibitory effects.
All eight affinity-matured antibodies displayed binding affinity towards SLC2A1 with varying
specificity levels, demonstrating their potential for targeted therapies. This study successfully
enhanced the affinity of antibody 6F12#33 through affinity maturation and phage display
techniques, providing valuable insights for the development of glucose metabolism-related cancer
treatments.
Description
Keywords
Phage display, affinity maturation, SLC2A1, monoclonal antibodies