Development of Improved High-Performance Liquid Chromatography Method for the Determination of Residual Caprylic Acid in Formulations of Human Immunoglobulins

Štimac, Adela; Kurtović, Tihana; Pavlović, Nediljko; Halassy, Beata

doi:10.3390/molecules27051665

Cited by 2 publications

(2 citation statements)

References 32 publications

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“…Prior work introduced DARQ as a simple mix-and-read assay for the quantification of therapeutic monoclonal antibodies Trastuzumab and Adalimumabrespectively used in fighting breast cancer and rheumatoid arthritisin bioprocess fluids . Our results showed that DARQ technology is endowed with numerous advantages compared to ELISA, BLI, and SPR assays (Table S1), − chiefly, high reproducibility, speed and ease of execution (mix-and-read), flexibility, robustness, and low cost of goods.…”

Section: Introductionmentioning

confidence: 86%

One-Step Quantification of anti-Covid-19 Antibodies via Dual Affinity Ratiometric Quenching Assays

et al. 2023

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The global pandemic caused by acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has affected millions of people and paralyzed healthcare systems worldwide. Developing rapid and accurate tests to detect and quantify anti-SARS-CoV-2 antibodies in complex fluids is critical to (i) track and address the spread of SARS-CoV-2 variants with different virulence and (ii) support the industrial manufacturing and clinical administration of anti-SARS-CoV-2 therapeutic antibodies. Conventional immunoassays, such as lateral flow, ELISA, and surface plasmon resonance (SPR), are either qualitative or, when quantitative, are laborious and expensive and suffer from high variability. Responding to these challenges, this study evaluates the performance of the Dual-Affinity Ratiometric Quenching (DARQ) assay for the quantification of anti-SARS-CoV-2 antibodies in bioprocess harvests and intermediate fractions (i.e., a Chinese hamster ovary (CHO) cell culture supernatant and a purified eluate) and human fluids (i.e., saliva and plasma). Monoclonal antibodies targeting the SARS-CoV-2 nucleocapsid as well as the spike protein of the delta and omicron variants are adopted as model analytes. Additionally, conjugate pads loaded with dried protein were studied as an at-line quantification method that can be used in clinical or manufacturing laboratories. Our results indicate that the DARQ assay is a highly reproducible (coefficient of variation ∼0.5–3%) and rapid (<10 min) test, whose sensitivity (∼0.23–2.5 ng/mL), limit of detection (23–250 ng/mL), and dynamic range (70–1300 ng/mL) are independent of sample complexity, thus representing a valuable tool for monitoring anti-SARS-CoV-2 antibodies.

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Section: Introductionmentioning

confidence: 86%

One-Step Quantification of anti-Covid-19 Antibodies via Dual Affinity Ratiometric Quenching Assays

et al. 2023

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“…Quantification of residual caprylic acid was determined by the validated HPLC method ( 26 ). Briefly, HPLC analysis was performed on RP C18 column (2.1 × 150 mm) (Shimadzu, Japan) with 0.05% TFA in water : ACN = 50 : 50 ( V / V ) as a mobile phase in isocratic mode at a flow rate of 0.2 mL min -1 on a Shimadzu HPLC system (Shimadzu, Japan).…”

Section: Methodsmentioning

confidence: 99%

Efficient and Sustainable Platform for Preparation of a High-Quality Immunoglobulin G as an Urgent Treatment Option During Emerging Virus Outbreaks

et al. 2022

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During the pre-vaccine era of the COVID-19 pandemic convalescent plasma has once again emerged as a major potential therapeutic form of passive immunization that in specific cases still represents irreplaceable treatment option. There is a growing concern that variable concentration of neutralizing antibodies, present in convalescent plasma which originates from different donors, apparently affects its effectiveness. The drawback can be overcome through the downstream process of immunoglobulin fraction purification into a standardized product of improved safety and efficacy. All modern procedures are quite lengthy processes. They are also based on fractionation of large plasma quantities whose collection is not attainable during an epidemic. When outbreaks of infectious diseases are occurring more frequently, there is a great need for a more sustainable production approach that would be goal-oriented towards assuring easily and readily available immunoglobulin of therapeutic relevance. We propose a refinement strategy for the IgG preparation achieved through simplification and reduction of the processing steps. It was designed as a small but scalable process to offer an immediately available treatment option that would simultaneously be harmonized with an increased availability of convalescent plasma over the viral outbreak time-course. Concerning the ongoing pandemic status of the COVID-19, the proof of concept was demonstrated on anti-SARS-CoV-2 convalescent plasma but is likely applicable to any other type depending on the current needs. It was guided by the idea of persistent keeping of IgG molecules in the solution, so that protection of their native structure could be assured. Our manufacturing procedure provided a high-quality IgG product of above the average recovery whose composition profile was analyzed by mass spectrometry as quality control check. It was proved free from IgA and IgM as mediators of adverse transfusion reactions, as well as of any other residual impurities, since only IgG fragments were identified. The proportion of S protein-specific IgGs remained unchanged relative to the convalescent plasma. Undisturbed IgG subclass composition was accomplished as well. However, the fractionation principle affected the final product’s capacity to neutralize wild-type SARS-CoV-2 infectivity, reducing it by half. Decrease in neutralization potency significantly correlated with the amount of IgM in the starting material.

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Development of Improved High-Performance Liquid Chromatography Method for the Determination of Residual Caprylic Acid in Formulations of Human Immunoglobulins

Cited by 2 publications

References 32 publications

One-Step Quantification of anti-Covid-19 Antibodies via Dual Affinity Ratiometric Quenching Assays

One-Step Quantification of anti-Covid-19 Antibodies via Dual Affinity Ratiometric Quenching Assays

Efficient and Sustainable Platform for Preparation of a High-Quality Immunoglobulin G as an Urgent Treatment Option During Emerging Virus Outbreaks

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