High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing

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Standard

High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing. / Nguyen, Jennifer Marie; Smith, Jacquelynn; Rzewuski, Susan; Legido-Quigley, Cristina; Lauber, Matthew A.

I: mAbs, Bind 11, Nr. 8, 2019, s. 1358-1366.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Nguyen, JM, Smith, J, Rzewuski, S, Legido-Quigley, C & Lauber, MA 2019, 'High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing', mAbs, bind 11, nr. 8, s. 1358-1366. https://doi.org/10.1080/19420862.2019.1658492

APA

Nguyen, J. M., Smith, J., Rzewuski, S., Legido-Quigley, C., & Lauber, M. A. (2019). High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing. mAbs, 11(8), 1358-1366. https://doi.org/10.1080/19420862.2019.1658492

Vancouver

Nguyen JM, Smith J, Rzewuski S, Legido-Quigley C, Lauber MA. High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing. mAbs. 2019;11(8):1358-1366. https://doi.org/10.1080/19420862.2019.1658492

Author

Nguyen, Jennifer Marie ; Smith, Jacquelynn ; Rzewuski, Susan ; Legido-Quigley, Cristina ; Lauber, Matthew A. / High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing. I: mAbs. 2019 ; Bind 11, Nr. 8. s. 1358-1366.

Bibtex

@article{a2546b731e2842aaada002fe2cce865f,
title = "High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing",
abstract = "Reversed-phase liquid chromatography (RPLC) separations of proteins using optical detection generally use trifluoroacetic acid (TFA) because it is a strong, hydrophobic acid and a very effective ion-pairing agent for minimizing chromatographic secondary interactions. Conversely and in order to avoid ion suppression, analyses entailing mass spectrometry (MS) detection is often performed with a weaker ion-pairing modifier, like formic acid (FA), but resolution quality may be reduced. To gain both the chromatographic advantages of TFA and the enhanced MS sensitivity of FA, we explored the use of an alternative acid, difluoroacetic acid (DFA). This acid modifier is less acidic and less hydrophobic than TFA and is believed to advantageously affect the surface tension of electrospray droplets. Thus, it is possible to increase MS sensitivity threefold by replacing TFA with DFA. Moreover, we have observed DFA ion pairing to concomitantly produce higher chromatographic resolution than FA and even TFA. For this reason, we prepared and used MS-quality DFA in place of FA and TFA in separations involving IdeS digested, reduced NIST mAb and a proprietary antibody-drug conjugate (ADC), aiming to increase sensitivity, resolution and protein recovery. The resulting method using DFA was qualified and applied to two other ADCs and gave heightened sensitivity, resolution and protein recovery versus analyses using TFA. This new method, based on a purified, trace metal free DFA, can potentially become a state-of-the-art liquid chromatography-MS technique for the deep characterization of ADCs.",
keywords = "Difluoroacetic acid, DFA, Formic acid, FA, Trifluoroacetic acid, TFA, Antibody-drug conjugate, ADC, IdeS digestion, Monoclonal antibody, mAb, NIST mAb, Reversed-phase chromatography, Subunit profiling, LC-MS, Peak capacity, Protein recovery, MS sensitivity, Disulfide isoforms, Drug-to-antibody ratio, DAR, Salt adducts, Metal adducts, Sodium, Potassium",
author = "Nguyen, {Jennifer Marie} and Jacquelynn Smith and Susan Rzewuski and Cristina Legido-Quigley and Lauber, {Matthew A}",
note = "CURIS 2019 NEXS 399",
year = "2019",
doi = "10.1080/19420862.2019.1658492",
language = "English",
volume = "11",
pages = "1358--1366",
journal = "mAbs",
issn = "1942-0862",
publisher = "Taylor & Francis",
number = "8",

}

RIS

TY - JOUR

T1 - High sensitivity LC-MS profiling of antibody-drug conjugates with difluoroacetic acid ion pairing

AU - Nguyen, Jennifer Marie

AU - Smith, Jacquelynn

AU - Rzewuski, Susan

AU - Legido-Quigley, Cristina

AU - Lauber, Matthew A

N1 - CURIS 2019 NEXS 399

PY - 2019

Y1 - 2019

N2 - Reversed-phase liquid chromatography (RPLC) separations of proteins using optical detection generally use trifluoroacetic acid (TFA) because it is a strong, hydrophobic acid and a very effective ion-pairing agent for minimizing chromatographic secondary interactions. Conversely and in order to avoid ion suppression, analyses entailing mass spectrometry (MS) detection is often performed with a weaker ion-pairing modifier, like formic acid (FA), but resolution quality may be reduced. To gain both the chromatographic advantages of TFA and the enhanced MS sensitivity of FA, we explored the use of an alternative acid, difluoroacetic acid (DFA). This acid modifier is less acidic and less hydrophobic than TFA and is believed to advantageously affect the surface tension of electrospray droplets. Thus, it is possible to increase MS sensitivity threefold by replacing TFA with DFA. Moreover, we have observed DFA ion pairing to concomitantly produce higher chromatographic resolution than FA and even TFA. For this reason, we prepared and used MS-quality DFA in place of FA and TFA in separations involving IdeS digested, reduced NIST mAb and a proprietary antibody-drug conjugate (ADC), aiming to increase sensitivity, resolution and protein recovery. The resulting method using DFA was qualified and applied to two other ADCs and gave heightened sensitivity, resolution and protein recovery versus analyses using TFA. This new method, based on a purified, trace metal free DFA, can potentially become a state-of-the-art liquid chromatography-MS technique for the deep characterization of ADCs.

AB - Reversed-phase liquid chromatography (RPLC) separations of proteins using optical detection generally use trifluoroacetic acid (TFA) because it is a strong, hydrophobic acid and a very effective ion-pairing agent for minimizing chromatographic secondary interactions. Conversely and in order to avoid ion suppression, analyses entailing mass spectrometry (MS) detection is often performed with a weaker ion-pairing modifier, like formic acid (FA), but resolution quality may be reduced. To gain both the chromatographic advantages of TFA and the enhanced MS sensitivity of FA, we explored the use of an alternative acid, difluoroacetic acid (DFA). This acid modifier is less acidic and less hydrophobic than TFA and is believed to advantageously affect the surface tension of electrospray droplets. Thus, it is possible to increase MS sensitivity threefold by replacing TFA with DFA. Moreover, we have observed DFA ion pairing to concomitantly produce higher chromatographic resolution than FA and even TFA. For this reason, we prepared and used MS-quality DFA in place of FA and TFA in separations involving IdeS digested, reduced NIST mAb and a proprietary antibody-drug conjugate (ADC), aiming to increase sensitivity, resolution and protein recovery. The resulting method using DFA was qualified and applied to two other ADCs and gave heightened sensitivity, resolution and protein recovery versus analyses using TFA. This new method, based on a purified, trace metal free DFA, can potentially become a state-of-the-art liquid chromatography-MS technique for the deep characterization of ADCs.

KW - Difluoroacetic acid

KW - DFA

KW - Formic acid

KW - FA

KW - Trifluoroacetic acid

KW - TFA

KW - Antibody-drug conjugate

KW - ADC

KW - IdeS digestion

KW - Monoclonal antibody

KW - mAb

KW - NIST mAb

KW - Reversed-phase chromatography

KW - Subunit profiling

KW - LC-MS

KW - Peak capacity

KW - Protein recovery

KW - MS sensitivity

KW - Disulfide isoforms

KW - Drug-to-antibody ratio

KW - DAR

KW - Salt adducts

KW - Metal adducts

KW - Sodium

KW - Potassium

U2 - 10.1080/19420862.2019.1658492

DO - 10.1080/19420862.2019.1658492

M3 - Journal article

C2 - 31500514

VL - 11

SP - 1358

EP - 1366

JO - mAbs

JF - mAbs

SN - 1942-0862

IS - 8

ER -

ID: 241218697