---
_id: '8237'
abstract:
- lang: eng
text: Monoclonal antibodies find broad application as therapy for various types
of cancer by employing multiple mechanisms of action against tumors. Manipulating
the Fc-mediated functions of antibodies that engage immune effector cells, such
as NK cells, represents a strategy to influence effector cell activation and to
enhance antibody potency and potentially efficacy. We developed a novel approach
to generate and ascertain the functional attributes of Fc mutant monoclonal antibodies.
This entailed coupling single expression vector (pVitro1) antibody cloning, using
polymerase incomplete primer extension (PIPE) polymerase chain reaction, together
with simultaneous Fc region point mutagenesis and high yield transient expression
in human mammalian cells. Employing this, we engineered wild type, low (N297Q,
NQ), and high (S239D/I332E, DE) FcR-binding Fc mutant monoclonal antibody panels
recognizing two cancer antigens, HER2/neu and chondroitin sulfate proteoglycan
4. Antibodies were generated with universal mutagenic primers applicable to any
IgG1 pVitro1 constructs, with high mutagenesis and transfection efficiency, in
small culture volumes, at high yields and within 12 days from design to purified
material. Antibody variants conserved their Fab-mediated recognition of target
antigens and their direct anti-proliferative effects against cancer cells. Fc
mutations had a significant impact on antibody interactions with Fc receptors
(FcRs) on human NK cells, and consequently on the potency of NK cell activation,
quantified by immune complex-mediated calcium mobilization and by antibody-dependent
cellular cytotoxicity (ADCC) of tumor cells. This strategy for manipulation and
testing of Fc region engagement with cognate FcRs can facilitate the design of
antibodies with defined effector functions and potentially enhanced efficacy against
tumor cells.
article_number: '1112'
article_processing_charge: No
article_type: original
author:
- first_name: Kristina M.
full_name: Ilieva, Kristina M.
last_name: Ilieva
- first_name: Judit
full_name: Fazekas-Singer, Judit
id: 36432834-F248-11E8-B48F-1D18A9856A87
last_name: Fazekas-Singer
orcid: 0000-0002-8777-3502
- first_name: Daniela Y.
full_name: Achkova, Daniela Y.
last_name: Achkova
- first_name: Tihomir S.
full_name: Dodev, Tihomir S.
last_name: Dodev
- first_name: Silvia
full_name: Mele, Silvia
last_name: Mele
- first_name: Silvia
full_name: Crescioli, Silvia
last_name: Crescioli
- first_name: Heather J.
full_name: Bax, Heather J.
last_name: Bax
- first_name: Anthony
full_name: Cheung, Anthony
last_name: Cheung
- first_name: Panagiotis
full_name: Karagiannis, Panagiotis
last_name: Karagiannis
- first_name: Isabel
full_name: Correa, Isabel
last_name: Correa
- first_name: Mariangela
full_name: Figini, Mariangela
last_name: Figini
- first_name: Rebecca
full_name: Marlow, Rebecca
last_name: Marlow
- first_name: Debra H.
full_name: Josephs, Debra H.
last_name: Josephs
- first_name: Andrew J.
full_name: Beavil, Andrew J.
last_name: Beavil
- first_name: John
full_name: Maher, John
last_name: Maher
- first_name: James F.
full_name: Spicer, James F.
last_name: Spicer
- first_name: Erika
full_name: Jensen-Jarolim, Erika
last_name: Jensen-Jarolim
- first_name: Andrew N.
full_name: Tutt, Andrew N.
last_name: Tutt
- first_name: Sophia N.
full_name: Karagiannis, Sophia N.
last_name: Karagiannis
citation:
ama: Ilieva KM, Singer J, Achkova DY, et al. Functionally active Fc mutant antibodies
recognizing cancer antigens generated rapidly at high yields. Frontiers in
Immunology. 2017;8. doi:10.3389/fimmu.2017.01112
apa: Ilieva, K. M., Singer, J., Achkova, D. Y., Dodev, T. S., Mele, S., Crescioli,
S., … Karagiannis, S. N. (2017). Functionally active Fc mutant antibodies recognizing
cancer antigens generated rapidly at high yields. Frontiers in Immunology.
Frontiers. https://doi.org/10.3389/fimmu.2017.01112
chicago: Ilieva, Kristina M., Judit Singer, Daniela Y. Achkova, Tihomir S. Dodev,
Silvia Mele, Silvia Crescioli, Heather J. Bax, et al. “Functionally Active Fc
Mutant Antibodies Recognizing Cancer Antigens Generated Rapidly at High Yields.”
Frontiers in Immunology. Frontiers, 2017. https://doi.org/10.3389/fimmu.2017.01112.
ieee: K. M. Ilieva et al., “Functionally active Fc mutant antibodies recognizing
cancer antigens generated rapidly at high yields,” Frontiers in Immunology,
vol. 8. Frontiers, 2017.
ista: Ilieva KM, Singer J, Achkova DY, Dodev TS, Mele S, Crescioli S, Bax HJ, Cheung
A, Karagiannis P, Correa I, Figini M, Marlow R, Josephs DH, Beavil AJ, Maher J,
Spicer JF, Jensen-Jarolim E, Tutt AN, Karagiannis SN. 2017. Functionally active
Fc mutant antibodies recognizing cancer antigens generated rapidly at high yields.
Frontiers in Immunology. 8, 1112.
mla: Ilieva, Kristina M., et al. “Functionally Active Fc Mutant Antibodies Recognizing
Cancer Antigens Generated Rapidly at High Yields.” Frontiers in Immunology,
vol. 8, 1112, Frontiers, 2017, doi:10.3389/fimmu.2017.01112.
short: K.M. Ilieva, J. Singer, D.Y. Achkova, T.S. Dodev, S. Mele, S. Crescioli,
H.J. Bax, A. Cheung, P. Karagiannis, I. Correa, M. Figini, R. Marlow, D.H. Josephs,
A.J. Beavil, J. Maher, J.F. Spicer, E. Jensen-Jarolim, A.N. Tutt, S.N. Karagiannis,
Frontiers in Immunology 8 (2017).
date_created: 2020-08-10T11:53:32Z
date_published: 2017-09-11T00:00:00Z
date_updated: 2021-01-12T08:17:39Z
day: '11'
doi: 10.3389/fimmu.2017.01112
extern: '1'
intvolume: ' 8'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://doi.org/10.3389/fimmu.2017.01112
month: '09'
oa: 1
oa_version: Published Version
publication: Frontiers in Immunology
publication_identifier:
issn:
- 1664-3224
publication_status: published
publisher: Frontiers
quality_controlled: '1'
status: public
title: Functionally active Fc mutant antibodies recognizing cancer antigens generated
rapidly at high yields
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 8
year: '2017'
...