Host cell protein testing strategy for hepatitis B antigen in Hexavalent vaccine – Towards a general testing strategy for recombinant vaccines

Toinon, Audrey; Fontaine, C.; Thion, Laurent; Gajewska, Beata; Carpick, Bruce; Nougarede, Nolwenn; Uhlrich, Sylvie

doi:10.1016/j.biologicals.2018.05.006

Cited by 7 publications

(3 citation statements)

References 7 publications

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“…ELISA is most commonly used to monitor HCPs during process development and ensure that acceptably low levels of HCPs are achieved [6]. However, there are several limitations with this technique.…”

Section: This Standard Based On Peptide Sequences Derived From 18mentioning

confidence: 99%

“…However, it is still essential to demonstrate that HCPs detected in the vaccine are present at low levels, that they do not represent any known safety risk to the vaccinated people, and that the HCPs profile is consistent from batch to batch. Therefore, pharmaceutical laboratories must assess HCPs levels and develop analytical tools for characterizing these impurities [6].…”

Section: Introductionmentioning

confidence: 99%

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An innovative standard for LC‐MS‐based HCP profiling and accurate quantity assessment: Application to batch consistency in viral vaccine samples

et al. 2021

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Biotherapeutics, molecules produced from biological systems, require rigorous purification steps to remove impurities including host cell proteins (HCPs). Regulatory guidelines require manufacturers to monitor process‐related impurities along the purification workflow. Mass spectrometry (MS) has recently been considered as a complementary method to the well‐established ELISA for HCPs quantification, since it has the advantage of unambiguously identifying individual HCP. In this study, we developed an innovative standard dedicated to MS‐based HCP profiling analysis in order to monitor the consistency of viral vaccine intermediate purification samples. This standard, termed the HCP‐PROFILER standard, is composed of a water‐soluble bead (READYBEADS technology) which, after being added into the sample, releases unlabeled peptides in controlled amounts. The standard meets three desired criteria: (1) it is composed of multiple peptides, at different concentration levels, allowing construction of a calibration curve covering the dynamic range of HCPs present in the target sample, ensuring quantification accuracy; (2) it demonstrates high batch‐to‐batch reproducibility, ensuring quantification robustness and consistency over time; and (3) it is easy to use and avoids user‐induced analytical biases. In this study, we present the use of the HCP‐PROFILER standard for vaccine batches comparison and downstream process performance studies.

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Section: This Standard Based On Peptide Sequences Derived From 18mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An innovative standard for LC‐MS‐based HCP profiling and accurate quantity assessment: Application to batch consistency in viral vaccine samples

et al. 2021

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“…In addition, it is also used in biomarker profiling [ 10 , 11 ] and biomarker-based diagnostic assays for cancer diagnosis [ 12 ], diagnosis of autoimmune diseases [ 13 ], and Alzheimer’s [ 14 ]. Moreover, it has applications in vaccine development [ 15 ] and conjugate vaccine assessment [ 16 ]. Also, it is of use in discrimination of allergic cultivates for human [ 17 ] and even as a basic bio-approach in engineering as in early-diagnostic biosensors [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?

Nadeem¹,

Barakat

Bahgat³

2021

Journal of Genetic Engineering and Biotechnology

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Background Sandwich ELISA is an ideal antigen detection and quantification assay. Recently, it was used as the basic concept for high technology diagnostics. The specificity of the assay depends on the exclusion of detection cross-reactivity which arises from using two antibodies developed in different species. Since mice and rats are the common laboratory animals used to develop antigen specific antibodies. Therefore, the questions we addressed here were (1) can one use antigen-specific antibodies raised in mice and rats in the same assay to specifically detect/quantify antigens? and (2) which antibodies of the two rodents should be placed for capturing and for detection in the antigen-detection sandwich? Results Direct ELISA assay was used to assess for the specific reaction of the HRP-conjugated antibody to the target serum. First reaction was to compare between either conjugate anti-rat IgG (homologous) or anti-mouse IgG (heterologous) for the detection of rat sera IgG. Following the dilution factor optimization, the O.D. were 0.744±0.051 and 0.604±0.05, respectively (p= .004). The difference in mean O.D. of 0.14 reflected an unaccepted non-specific reaction. The second reaction was to compare between either conjugate anti-mouse IgG (homologous) and anti-rat IgG (heterologous) for the detection of mouse sera IgG. The recorded O.D. were 0.9414±0.14 and 0.317 ±0.141, respectively (p= .0002). The improved difference in mean O.D. of 0.624 reflecting a minimized cross-reaction. Conclusions Our results suggest that it is possible to use both Swiss albino mice and albino rats in a single sandwich ELISA, given that the captured antibody species to be from the Swiss albino mice and the detection antibody to be from the albino rat. The described working details are limited to the source of the antibodies used in the study. However, the approach stresses on the importance of such optimization steps before making any interpretations based on the antigen detection. To our knowledge, this study is the first to cover the optimal order of the capturing and the detection antibodies in a sandwich ELISA assay. In addition to addressing the possible interfering cross-reactivity that result from using mouse and rat serum antibodies in a single assay. Graphical abstract

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Recent applications of quantitative mass spectrometry in biopharmaceutical process development and manufacturing

2023

Journal of Pharmaceutical and Biomedical Analysis

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Host cell protein testing strategy for hepatitis B antigen in Hexavalent vaccine – Towards a general testing strategy for recombinant vaccines

Cited by 7 publications

References 7 publications

An innovative standard for LC‐MS‐based HCP profiling and accurate quantity assessment: Application to batch consistency in viral vaccine samples

An innovative standard for LC‐MS‐based HCP profiling and accurate quantity assessment: Application to batch consistency in viral vaccine samples

Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?

Recent applications of quantitative mass spectrometry in biopharmaceutical process development and manufacturing

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