Antibody-based biotherapeutics are the fastest growing category of therapeutics
entering clinical studies [1]. The popularity is attributed to the high selectivity of
these drugs which enhances their efficacy and reduces systemic toxicity, in turn,
increasing the therapeutic index. In addition, antibody-based biotherapeutics have
long circulatory half-lives and are less likely to undergo significant biotransformation
in vivo. Since the approval of the first monoclonal antibody (mAb) drug (Orthoclone
OKT3®) in the 1980s, the number of antibody-based biotherapeutics on the mar-
ket and in development has reached several hundred. Recent advancements in
antibody engineering and manufacturing techniques have led to generation of new
biotherapeutic modalities such as multispecific mAbs, antibody-drug conjugates,
Probody™ therapeutics and Probody™ drug conjugates, PEGylated antibody frag-
ments and fragment crystallizable (Fc)-fusion proteins. The increasing complexity
of the new biotherapeutic modalities pose newer bioanalytical challenges. In some
instances, multiple bioanalytical assays using different analytical platforms are re-
quired to address the pharmacokinetic characterization of these new modalities.
Bioanalysis by LC-MS for Antibody-Based Biotherapeutics
Traditionally, ligand binding assays (LBA), primarily enzyme-linked immunosorbent
assay (ELISA), have been the gold standard for protein quantification. ELISA has
numerous benefits such as high-sensitivity, high-throughput, low-cost and ease of
use. LBAs require high-quality critical reagents, that have high selectivity for the
biotherapeutic drug, for capture and detection. The availability of a highly selective
critical reagents is a major challenge of LBAs. The time of production of a high-qual-
ity critical reagent can prolong the method development time. The narrow dynamic
range (typically around 100-fold) and high susceptibility to matrix interferences are
other limitations of the LBAs.
Over past decade, liquid chromatography and mass spectrometry (LC-MS) has
emerged as an alternate tool for quantification of proteins [2]. Rapid method de-
velopment, broader dynamic range, higher selectivity and multiplexing capabilities
are the main advantages of the LC-MS platform in comparison to the LBAs. The
advancements in the mass spectrometric instrumentation and sample preparation
methodology has enabled the use LC-MS as a complementary bioanalytical tool
and in some applications as a mainstream bioanalytical tool for protein biothera-
peutics.
Quantitative bioanalysis of antibody-based biotherapeutics by LC-MS is typically
PPD Laboratories, Richmond, VA 23230, USA.
Correspondence: PPD Laboratories, 2244 Dabney Road, Richmond, VA 23230, USA. Phone:
+1 804 977 8491; Email: morse.faria@ppdi.com
Addressing Bioanalytical Needs of Antibody-Based Biotherapeutics
by LC-MS
Morse Faria
FOREWORD
Citation:
Faria M. Addressing Bioanalytical
Needs of Antibody-Based Biothera-
peutics by LC-MS.
J Appl Bioanal 6(1), 7-11 (2020).
Open Access & Copyright:
©2020 Faria M. This is an open
access article distributed under the
terms of the Creative Commons Attri-
bution License (CC-BY) which permits
any use, distribution, and reproduction
in any medium, provided the original
author(s) and source are credited.
Funding & Manuscript writing
assistance:
The author has declared that no fund-
ing nor writing assistance was utilized
in the production of this article.
Financial & Competing interests:
The author is employee of PPD Labo-
ratories USA. The author has declared
that no competing interest exist.
Journal of
Applied Bioanalysis
JOURNAL OF APPLIED BIOANALYSIS Volume 6, No. 1 | January 2020 7
http://dx.doi.org/10.17145/jab.20.003
openaccess