Preferential germline usage and VH/VL pairing observed in human antibodies selected by mRNA display.
Since the invention of phage display, in vitro antibody display technologies have revolutionized the field of antibody discovery. In combination with antibody libraries constructed with sequences of human origin, such technologies enable accelerated therapeutic antibody discovery while bypassing the laborious animal immunization and hybridoma generation processes. Many in vitro display technologies developed since aim to differentiate from phage display by displaying full-length IgG proteins, utilizing eukaryotic translation system and codons, increasing library size or real-time kinetic selection by fluorescent activated cell sorting. We report here the development of an mRNA display technology and an accompanying HCDR3 size spectratyping monitor for human antibody discovery. Importantly, the mRNA display technology maintains a monovalent linkage between the mRNA (genotype) and display binding protein (phenotype), which minimizes avidity effect common in other display systems and allows for a stringent affinity and off-rate selection. The mRNA display technology successfully identified 100 human antibodies in 15 different selections against various targets from naïve human antibody libraries. These antibodies in general have high affinity and diversity. By analyzing the germline usage and combination of antibodies selected by the mRNA display technology, we identified trends and determined the productivity of each germline subgroup in the libraries that could serve as the knowledge base for constructing fully synthetic, next generation antibody libraries.
AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
Lei
L
Chen
AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
Yuliya A
YA
Kutskova
AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
Feng
F
Hong
AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
John E
JE
Memmott
AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
Suju
S
Zhong
AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
Megan D
MD
Jenkinson
AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA chung-ming.hsieh@abbvie.com.
Chung-Ming
CM
Hsieh
D061505:Cell Surface Display Techniques / Q000379:methods*; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics; D061905:Single-Domain Antibodies / Q000235:genetics*
D012333:RNA, Messenger; D061905:Single-Domain Antibodies
fully human antibody; germline pairing; germline usage; mRNA display; spectratyping