Identification of New Antibodies Targeting Malignant Plasma Cells for Immunotherapy by Next-Generation Sequencing-Assisted Phage Display
Standard
Identification of New Antibodies Targeting Malignant Plasma Cells for Immunotherapy by Next-Generation Sequencing-Assisted Phage Display. / Krohn, Steffen; Boje, Ammelie Svea; Gehlert, Carina Lynn; Lutz, Sebastian; Darzentas, Nikos; Knecht, Henrik; Herrmann, Dietrich; Brüggemann, Monika; Scheidig, Axel J; Weisel, Katja; Gramatzki, Martin; Peipp, Matthias; Klausz, Katja.
In: FRONT IMMUNOL, Vol. 13, 908093, 2022.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Identification of New Antibodies Targeting Malignant Plasma Cells for Immunotherapy by Next-Generation Sequencing-Assisted Phage Display
AU - Krohn, Steffen
AU - Boje, Ammelie Svea
AU - Gehlert, Carina Lynn
AU - Lutz, Sebastian
AU - Darzentas, Nikos
AU - Knecht, Henrik
AU - Herrmann, Dietrich
AU - Brüggemann, Monika
AU - Scheidig, Axel J
AU - Weisel, Katja
AU - Gramatzki, Martin
AU - Peipp, Matthias
AU - Klausz, Katja
N1 - Copyright © 2022 Krohn, Boje, Gehlert, Lutz, Darzentas, Knecht, Herrmann, Brüggemann, Scheidig, Weisel, Gramatzki, Peipp and Klausz.
PY - 2022
Y1 - 2022
N2 - To identify new antibodies for the treatment of plasma cell disorders including multiple myeloma (MM), a single-chain Fragment variable (scFv) antibody library was generated by immunizing mice with patient-derived malignant plasma cells. To enrich antibodies binding myeloma antigens, phage display with cellular panning was performed. After depleting the immune library with leukocytes of healthy donors, selection of antibodies was done with L-363 plasma cell line in two consecutive panning rounds. Monitoring the antibodies' enrichment throughout the panning by next-generation sequencing (NGS) identified several promising candidates. Initially, 41 unique scFv antibodies evolving from different B cell clones were selected. Nine of these antibodies strongly binding to myeloma cells and weakly binding to peripheral blood mononuclear cells (PBMC) were characterized. Using stably transfected Chinese hamster ovary cells expressing individual myeloma-associated antigens revealed that two antibodies bind CD38 and intercellular adhesion molecule-1 (ICAM-1), respectively, and 7 antibodies target yet unknown antigens. To evaluate the therapeutic potential of our new antibodies, in a first proof-of-concept study the CD38 binding scFv phage antibody was converted into a chimeric IgG1. Further analyses revealed that #5-CD38-IgG1 shared an overlapping epitope with daratumumab and isatuximab and had potent anti-myeloma activity comparable to the two clinically approved CD38 antibodies. These results indicate that by phage display and deep sequencing, new antibodies with therapeutic potential for MM immunotherapy can be identified.
AB - To identify new antibodies for the treatment of plasma cell disorders including multiple myeloma (MM), a single-chain Fragment variable (scFv) antibody library was generated by immunizing mice with patient-derived malignant plasma cells. To enrich antibodies binding myeloma antigens, phage display with cellular panning was performed. After depleting the immune library with leukocytes of healthy donors, selection of antibodies was done with L-363 plasma cell line in two consecutive panning rounds. Monitoring the antibodies' enrichment throughout the panning by next-generation sequencing (NGS) identified several promising candidates. Initially, 41 unique scFv antibodies evolving from different B cell clones were selected. Nine of these antibodies strongly binding to myeloma cells and weakly binding to peripheral blood mononuclear cells (PBMC) were characterized. Using stably transfected Chinese hamster ovary cells expressing individual myeloma-associated antigens revealed that two antibodies bind CD38 and intercellular adhesion molecule-1 (ICAM-1), respectively, and 7 antibodies target yet unknown antigens. To evaluate the therapeutic potential of our new antibodies, in a first proof-of-concept study the CD38 binding scFv phage antibody was converted into a chimeric IgG1. Further analyses revealed that #5-CD38-IgG1 shared an overlapping epitope with daratumumab and isatuximab and had potent anti-myeloma activity comparable to the two clinically approved CD38 antibodies. These results indicate that by phage display and deep sequencing, new antibodies with therapeutic potential for MM immunotherapy can be identified.
KW - Animals
KW - Bacteriophages
KW - CHO Cells
KW - Cricetinae
KW - Cricetulus
KW - High-Throughput Nucleotide Sequencing
KW - Immunoglobulin G
KW - Immunologic Factors
KW - Immunotherapy
KW - Leukocytes, Mononuclear
KW - Mice
KW - Peptide Library
KW - Plasma Cells
U2 - 10.3389/fimmu.2022.908093
DO - 10.3389/fimmu.2022.908093
M3 - SCORING: Journal article
C2 - 35784366
VL - 13
JO - FRONT IMMUNOL
JF - FRONT IMMUNOL
SN - 1664-3224
M1 - 908093
ER -
