Investigating the utility of minimized sample preparation and high-resolution mass spectrometry for quantification of monoclonal antibody drugs

Nguyen, Tam T. T. N.; Mistarz, Ulrik H.; Costa, Narciso; Herbet, Amaury; Boquet, Didier; Bécher, François; Rand, Kasper D.

doi:10.1016/j.jpba.2018.07.012

Cited by 8 publications

(11 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A stable isotope-labeled (SIL)peptide of the signature peptide is most commonly used as an internal standard, which has been widely used in quantitative proteomics. However, there is no clear consensus regarding when the SIL-peptide should be added; the SIL-peptide was reported to be added before sample preparation 33,34 , before reduction 35,36 , before digestion 37,38 , or after digestion 39,40 .…”

Section: Discussionmentioning

confidence: 99%

“…A stable isotope-labeled (SIL)-peptide of the signature peptide is most commonly used as an internal standard, which has been widely used in quantitative proteomics. However, there is no clear consensus regarding when the SIL-peptide should be added; the SIL-peptide was reported to be added before sample preparation 33,34 , before reduction 35,36 , before digestion 37,38 , or after digestion 39,40 . Moreover, the use of SIL-peptide may not correct for variations in LC-MS detection especially during steps before mAb digestion as the physical and chemical behavior of the signature peptide within the intact mAb and the SIL-peptide can be different.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

Iwamoto

Koguchi

Yokoyama

et al. 2022

Preprint

View full text Add to dashboard Cite

Accurate quantitation of antibody is critical for development of monoclonal antibody therapeutics (mAbs). Therapeutic drug monitoring has been applied to measure levels of mAbs in clinics for dose adjustment for autoimmune disease. Trough levels of mAbs can be a biomarker for cancer immunotherapy. Thus, the deployment of a rapid and universal platform for mAb monitoring may benefit processes ranging from drug development to clinical practice for a wide spectrum of diseases. However, mAb monitoring often requires development and conduct of an individual ligand binding assay such as ELISA, which is impractical to scale. We streamlined quantitation of antibody therapeutics by a nano-surface and molecular-orientation limited (nSMOL) proteolysis assay using LC-MS with a universal reference antibody (refmAb-Q), for accurate multiplexed quantitation of unique signature peptides derived from mAbs. This innovative refmAb-Q nSMOL platform may provide a practical solution for quantitating an ever-increasing number of mAbs from developmental to clinical use settings.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

Iwamoto

Koguchi

Yokoyama

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…The sample preparation is influenced by matrix effect from lipids, proteins, and salts and the protease must be optimized to the total protein concentration in the sample for optimal digestion efficiency [17,18]. This method is applied on a routine basis for mAb in plasma samples [19][20][21][22][23], and though CSF appears as a cleaner matrix with less proteins and lipids (Table 1), some key differences must be noted, namely the low sample volume and complicated sample collection. While approximately 15 mL CSF can be collected from humans during a single sampling event [24], the situation is much different for animal species commonly used in preclinical drug development.…”

Section: Introductionmentioning

confidence: 99%

Investigating surrogate cerebrospinal fluid matrix compositions for use in quantitative LC-MS analysis of therapeutic antibodies in the cerebrospinal fluid

et al. 2020

Self Cite

View full text Add to dashboard Cite

As quantitative analysis of biotherapeutics in cerebrospinal fluid (CSF) with LC-MS becomes increasingly widespread, there is a need for method developments towards higher sensitivity. By using artificial CSF (aCSF) in the development phase, the consumption of costly and sparsely available CSF can be limited. The aCSF compositions tested here were made from various dilutions of bovine serum albumin (BSA) or rat plasma to mimic the total protein concentration found in CSF. Focusing on monoclonal antibodies, the aCSF was spiked with human immunoglobulin (hIgG) and prepared with the bottom-up analysis technique using LC-MS. Assuming that the composition of the aCSF would affect the digest, the response from aCSF matrices was compared with CSF from rat, monkey, and dog in terms of estimated sample concentration and matrix effects. The samples were spiked with hIgG in the range of 10 to 1000 ng/mL and volumes of 10 μL were transferred to sample preparation. The results indicate that BSA dilutions from 300 to 2000 μg/mL and rat plasma dilutions of 0.5-2% provide the most accurate concentration estimates when compared with rat CSF. 1000 μg/mL BSA did not produce significantly different concentration estimates for 500 ng/mL samples when compared with CSF from rat, monkey, and dog, and can therefore be used as aCSF for several different species.

show abstract

“…In general, the development of human mAb therapeutics often requires robust mouse models for early stage screening and evaluation. , More recently, assessing pharmacokinetics (PK) and target mediated clearance as well as understanding PK/PD/E relationship of surrogate mouse mAb (a mouse antibody against the mouse counterpart of a human target) in immunocompetent mice have been deemed highly valuable in the translation of preclinical results and predicting human pharmacologically active dose and advancing discovery programs. , Accordingly, the demand of measuring such surrogates in circulation is expected to rise over time. However, the limited differences between surrogates and endogenous mouse immunoglobulins pose major challenges to the bioanalytical community because highly specific reagents, such as target antigens or anti-idiotypic antibodies, may not be readily available in the early discovery stage.…”

mentioning

confidence: 99%

“…Unlike LBA, if a unique surrogate peptide with good ionization efficiency and selectivity can be identified from the surrogate antibody (mouse counterpart) sequence, pellet digestion coupled with high resolution mass spectrometry 18 or immunoaffinity LC/MS/MS of unit resolution may provide another opportunity to distinguish analyte responses from the high immunoglobulin background. Immunoaffinity enrichment using protein A/G or antiframework antibodies can be viable options due to the better sample cleanup and potentially improved sensitivity compared to the pellet digestion.…”

mentioning

confidence: 99%

An Automated Multicycle Immunoaffinity Enrichment Approach Developed for Sensitive Mouse IgG1 Antibody Drug Analysis in Mouse Plasma Using LC/MS/MS

Bebrin

Piatkov

et al. 2021

Anal. Chem.

View full text Add to dashboard Cite

In the immuno-oncology field, surrogate mouse monoclonal antibodies are often preferred in establishing proper PK/PD/efficacy correlations as well as supporting anticipated mouse to human translation. Thus, a highly sensitive and specific bioanalytical method is needed in quantifying those surrogate mouse antibodies after dosing in mice. Unfortunately, when specific reagents, such as recombinant target antigen and anti-idiotypic antibody, are not available, measuring mouse surrogate antibody drugs in mice is very challenging for ligand binding assay (LBA) due to the severe cross reactivity potential. Different from LBA, if at least one unique surrogate peptide can be identified from the surrogate antibody sequence, the immunoaffinity enrichment based LC/MS/MS assay may be able to differentiate the analyte response from the high endogenous immunoglobulin background and provide adequate sensitivity. Herein, a new automated multicycle immunoaffinity enrichment method was recently developed to extract a surrogate mouse IgG1 (mIgG1) antibody drug from mouse plasma using a commercially available antimouse IgG1 secondary antibody. In the assay, reuse of the capture antibody up to six times mostly resolved the binding capacity issue caused by the abundant endogenous mIgG1 and made the immunoaffinity enrichment step more cost-effective. Combined with a unique surrogate peptide identified from the antibody, the LC/MS/MS assay achieved a limit of quantitation of 5 ng/mL with satisfactory assay precision, accuracy, and dynamic range. The successful implementation of this novel approach in discovery pharmacokinetic (PK) studies eliminates the dependence on specially generated immunoaffinity capturing reagents.

show abstract

Investigating the utility of minimized sample preparation and high-resolution mass spectrometry for quantification of monoclonal antibody drugs

Cited by 8 publications

References 35 publications

A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

A rapid and universal liquid chromatograph-mass spectrometry-based platform, refmAb-Q nSMOL, for monitoring monoclonal antibody therapeutics

Investigating surrogate cerebrospinal fluid matrix compositions for use in quantitative LC-MS analysis of therapeutic antibodies in the cerebrospinal fluid

An Automated Multicycle Immunoaffinity Enrichment Approach Developed for Sensitive Mouse IgG1 Antibody Drug Analysis in Mouse Plasma Using LC/MS/MS

Contact Info

Product

Resources

About