Nasal Epithelial Lining Fluid (NELF) Assessment of SARS-Cov-2 Vaccine-Induced Antibody Production

Rebuli, Meghan E.; Cao, K.T.; Cobos-Uribe, Catalina; Knight, Noelle; Jaspers, Ilona

doi:10.1164/ajrccm-conference.2022.205.1_meetingabstracts.a1691

Cited by 1 publication

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“…However, analyses of drug concentrations in NLF have been hindered by a lack of convenient, reliable sampling methods (e.g., swabs or nasal lavage) that yield low analyte concentrations which, in turn, produce broad reported ranges for the partition ratio of biopharmaceuticals into the nasopharynx. Common methods for mAb bioanalysis in NLF consist of ELISA, ,− electrochemiluminescence, , bead-based antibody assays, , and other commercially available serological assays . Generally, the ELISA assays were semiquantitative, , intended for research purposes only (with manufacturer reported range of 250–16,000 pg/mL), or reported standard curve in the ng/mL range (0.195–200 ng/mL) .…”

Section: Discussionmentioning

confidence: 99%

Multiplex Hybrid Antigen-Capture LC-MRM Quantification in Sera and Nasal Lining Fluid of AZD7442, a SARS-CoV-2-Targeting Antibody Combination

Huang

Bouquet

et al. 2022

Anal. Chem.

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AZD7442 (tixagevimab [AZD8895]/cilgavimab [AZD1061]) is a monoclonal antibody (mAb) combination in development for the prevention and treatment of coronavirus disease 2019. Traditionally, bioanalysis of mAbs is performed using ligand binding assays (LBAs), which offer sensitivity, robustness, and ease of implementation. However, LBAs frequently require generation of critical reagents that typically take several months. Instead, we developed a highly sensitive (5 ng/mL limit of quantification) method using a hybrid LBA-liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) approach for quantification of the two codosed antibodies in serum and nasal lining fluid (NLF), a rare matrix. The method was optimized by careful selection of multiple reaction monitoring, capture reagents, magnetic beads, chromatographic conditions, evaluations of selectivity, and matrix effect. The final assay used viral spike protein receptor-binding domain as capture reagent and signature proteotypic peptides from the complementarity-determining region of each mAb for detection. In contrast to other methods of similar/superior sensitivity, our approach did not require multidimensional separations and can be operated in an analytical flow regime, ensuring high throughput and robustness required for clinical analysis at scale. The sensitivity of this method significantly exceeds typical sensitivity of ∼100 ng/mL for analytical flow 1D LBA-LC-MS/MS methods for large macromolecules, such as antibodies. Furthermore, infection and vaccination status did not impact method performance, ensuring method robustness and applicability to a broad patient population. This report demonstrated the general applicability of the hybrid LBA-LC-MS/MS approach to platform quantification of antibodies with high sensitivity and reproducibility, with specialized extension to matrices of increasing interest, such as NLF.

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