PMID- 30880262
OWN - NLM
STAT- MEDLINE
DCOM- 20200427
LR  - 20200427
IS  - 1872-7905 (Electronic)
IS  - 0022-1759 (Linking)
VI  - 468
DP  - 2019 May
TI  - A novel method for determining antibody-dependent cellular phagocytosis.
PG  - 55-60
LID - S0022-1759(18)30119-4 [pii]
LID - 10.1016/j.jim.2019.03.001 [doi]
AB  - Antibody-based therapeutics are powerful tools to treat disease. While their 
      mechanism of action (MOA) always involves binding to a specific target via the 
      Fab region of the antibody, the induction of effector functions through the Fc 
      region of the antibody is equally important for antibody therapeutics designed to 
      deplete tumor cells. By binding of the Fc region to Fc gamma receptors (FcgammaRs) on 
      the surface of immune cells or complement factors, antibody therapeutics exert 
      effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and 
      complement-dependent cytotoxicity (CDC), both of which induce target cell death 
      and aid in the efficacy of treatment. Another major Fc effector function is 
      antibody-dependent cellular phagocytosis (ADCP). ADCP is the mechanism by which 
      antibody-opsonized target cells activate the FcgammaRs on the surface of macrophages 
      to induce phagocytosis, resulting in internalization and degradation of the 
      target cell through phagosome acidification. ADCP has been implicated as a major 
      MOA of several biologics, but this activity is difficult to measure in in vitro. 
      Most assays measure the association of target cells and macrophages; however, 
      co-localization can represent cell attachment rather than internalization. Here, 
      we describe the development of a novel method to accurately measure ADCP 
      activity. By labeling target cells with a pH sensitive dye that only fluoresces 
      in mature phagosomes, the ADCP activity of antibody therapeutics can be 
      accurately quantitated via flow cytometry.
CI  - Copyright (c) 2019 Elsevier B.V. All rights reserved.
FAU - Kamen, Lynn
AU  - Kamen L
AD  - Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San 
      Francisco, CA 94080-4990, United States.
FAU - Myneni, Srividya
AU  - Myneni S
AD  - Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San 
      Francisco, CA 94080-4990, United States.
FAU - Langsdorf, Chris
AU  - Langsdorf C
AD  - Thermo Fisher Scientific, 29851 Willow Creek Road, Eugene, OR 94702, United 
      States.
FAU - Kho, Elviza
AU  - Kho E
AD  - Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San 
      Francisco, CA 94080-4990, United States.
FAU - Ordonia, Benjamin
AU  - Ordonia B
AD  - Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San 
      Francisco, CA 94080-4990, United States.
FAU - Thakurta, Tara
AU  - Thakurta T
AD  - Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San 
      Francisco, CA 94080-4990, United States.
FAU - Zheng, Kai
AU  - Zheng K
AD  - Department of Late Stage Pharmaceutical Development, Genentech Inc, 1 DNA Way, 
      South San Francisco, CA 94080-4990, United States.
FAU - Song, An
AU  - Song A
AD  - Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San 
      Francisco, CA 94080-4990, United States; Immune-Onc Therapeutics, 4030 Fabian 
      Way, Palo Alto, CA 94303, United States.
FAU - Chung, Shan
AU  - Chung S
AD  - Department of BioAnalytical Sciences, Genentech Inc, 1 DNA Way, South San 
      Francisco, CA 94080-4990, United States. Electronic address: chung.shan@gene.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190314
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Antibodies, Monoclonal, Humanized)
RN  - 0 (Antineoplastic Agents, Immunological)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Receptors, IgG)
RN  - 4F4X42SYQ6 (Rituximab)
RN  - A10SJL62JY (ocrelizumab)
SB  - IM
MH  - Antibodies, Monoclonal, Humanized/metabolism/*pharmacology
MH  - Antineoplastic Agents, Immunological/*pharmacology
MH  - Cell Line, Tumor
MH  - Cytotoxicity, Immunologic/*drug effects
MH  - *Flow Cytometry
MH  - Fluorescent Dyes/*metabolism
MH  - Glycosylation
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Macrophages/*drug effects/immunology/metabolism/pathology
MH  - Neoplasms/*drug therapy/immunology/metabolism/pathology
MH  - Phagocytosis/*drug effects
MH  - Phagosomes/*drug effects/immunology/metabolism/pathology
MH  - Receptors, IgG/metabolism
MH  - Rituximab/metabolism/*pharmacology
OTO - NOTNLM
OT  - Antibody-dependent cellular phagocytosis
OT  - Cell-based assay
OT  - Flow cytometry
OT  - Glycosylation variants
OT  - pHrodo green AM
EDAT- 2019/03/19 06:00
MHDA- 2020/04/28 06:00
CRDT- 2019/03/19 06:00
PHST- 2018/03/23 00:00 [received]
PHST- 2019/03/12 00:00 [revised]
PHST- 2019/03/13 00:00 [accepted]
PHST- 2019/03/19 06:00 [pubmed]
PHST- 2020/04/28 06:00 [medline]
PHST- 2019/03/19 06:00 [entrez]
AID - S0022-1759(18)30119-4 [pii]
AID - 10.1016/j.jim.2019.03.001 [doi]
PST - ppublish
SO  - J Immunol Methods. 2019 May;468:55-60. doi: 10.1016/j.jim.2019.03.001. Epub 2019 
      Mar 14.