Fast and highly selective determination of hemagglutinin content in quadrivalent influenza vaccine by reversed-phase high-performance liquid chromatography method

Lorbetskie, Barry; Fortin, Nathalie; Durno, Laura; Wang, Junzhi; Li, Changgui; Li, Xuguang; Girard, Michel; Sauvé, Simon

doi:10.1016/j.chroma.2017.10.041

Cited by 6 publications

(14 citation statements)

References 28 publications

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“…Prior to RP-HPLC analysis, both the ncHA and cHA fractions were treated with DTT to cleave disulfide bonds between HA1 and HA2 moieties, as reported elsewhere. 29,38 Typically, the sum of HA1 peak areas from the cHA and ncHA fractions was found to be very close to that of HA1 peak areas obtained when no pretreatment was carried out.…”

Section: ■ Results and Discussionmentioning

confidence: 57%

“…35,36 Development of a sialic acid bearing receptor-based strategy could ensure that the RBS of trimeric HA found in the vaccine preparation is in a conformation supporting immunization response from individuals toward predicted antigenic strains. 35−37 Recently, updated RP-HPLC methods capable of separating and quantifying HA strains in trivalent 29 and in quadrivalent influenza vaccines, 38 the latter containing two A and two B strains, were reported. As mentioned above, such an approach is limited by its inability to distinguish between well-folded and stressed HA material and, as such, may not be directly used for potency determination.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Briefly, chromatographic separations were carried out at 55 °C with an AB gradient elution of 19 min at a flow rate of 1.0 mL/min where the composition of eluent A was 0.04% (v/v) aqueous TFA and eluent B was 0.03% (v/v) TFA in 25% ACN and 75% 2-propanol. 38 Separation conditions to measure BR binding and NA inhibition was performed on the Zorbax 300SB-C18, 150 mm × 4.6 mm, 3.5 μm particles (Agilent, Santa Clara, CA, USA). Briefly, chromatographic separations were carried out at 25 °C with an AB gradient elution of 20 min at a flow rate of 1.0 mL/min where the composition of eluent A was 50 mM potassium phosphate monobasic, pH 2.5, and eluent B was ACN.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Recently, updated RP-HPLC methods capable of separating and quantifying HA strains in trivalent and in quadrivalent influenza vaccines, the latter containing two A and two B strains, were reported. As mentioned above, such an approach is limited by its inability to distinguish between well-folded and stressed HA material and, as such, may not be directly used for potency determination.…”

Section: Introductionmentioning

confidence: 99%

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Selective Capture and Determination of Receptor-Binding Hemagglutinin in Influenza Vaccine Preparations Using a Coupled Receptor-Binding/RP-HPLC Assay

et al. 2019

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Influenza vaccine potency is determined by the quantification of immunologically active hemagglutinin capable of eliciting neutralizing antibodies upon immunization. Currently, the single radial immunodiffusion (SRID) method is the standard in vitro potency assay used for lot release of seasonal inactivated influenza vaccines. Despite the proven usage of SRID, significant limitations such as the timeconsuming preparation of reagents and limited dynamic range warrant the need for the development of alternative potency assays. Such alternative approaches need to discriminate and quantify relevant hemagglutinin material, provide strain identity, and be independent of strain-specific and seasonal reagents. Herein, we present a proof of concept method that combines the capture of conformationally well-folded hemagglutinin via a sialic acid binding step with the resolving power of reversed-phase high-performance liquid chromatography for strain identity and determination. Details of the protocol for the selective capture of receptor-binding hemagglutinin, its release from the receptor, and its relative determination are presented. This approach was found to provide flexibility for the reagents to be used and was adaptable to varying strain compositions of influenza vaccines. This proof of concept approach was developed as an antibody-independent methodology.

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Section: ■ Results and Discussionmentioning

confidence: 57%

Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Selective Capture and Determination of Receptor-Binding Hemagglutinin in Influenza Vaccine Preparations Using a Coupled Receptor-Binding/RP-HPLC Assay

et al. 2019

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“…More recently, a fast and highly selective method to determine HA content in quadrivalent influenza vaccine by reversed‐phase HPLC was reported. This HPLC method showed excellent resolution of all four hemagglutinins and is also the only physicochemical method capable of distinguishing the B strains in quadrivalent influenza vaccines . The eluents used for this HPLC method were 0.04% (v/v) aqueous trifluoroacetic acid (TFA) and 0.03% (v/v) TFA in 25% ACN and 75% 2‐propanol.…”

Section: Role Of Mass Spectrometry In the Vaccine Developmentmentioning

confidence: 99%

The expanding role of mass spectrometry in the field of vaccine development

Sharma

Sharma²,

Glick

2018

Mass Spectrometry Reviews

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Biological mass spectrometry has evolved as a core analytical technology in the last decade mainly because of its unparalleled ability to perform qualitative as well as quantitative profiling of enormously complex biological samples with high mass accuracy, sensitivity, selectivity and specificity. Mass spectrometry-based techniques are also routinely used to assess glycosylation and other post-translational modifications, disulfide bond linkage, and scrambling as well as for the detection of host cell protein contaminants in the field of biopharmaceuticals. The role of mass spectrometry in vaccine development has been very limited but is now expanding as the landscape of global vaccine development is shifting towards the development of recombinant vaccines. In this review, the role of mass spectrometry in vaccine development is presented, some of the ongoing efforts to develop vaccines for diseases with global unmet medical need are discussed and the regulatory challenges of implementing mass spectrometry techniques in a quality control laboratory setting are highlighted.

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Selective reversed-phase high-performance liquid chromatography method for the determination of intact SARS-CoV-2 spike protein

Lorbetskie

White

Creskey

et al. 2022

Journal of Chromatography A

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Fast and highly selective determination of hemagglutinin content in quadrivalent influenza vaccine by reversed-phase high-performance liquid chromatography method

Cited by 6 publications

References 28 publications

Selective Capture and Determination of Receptor-Binding Hemagglutinin in Influenza Vaccine Preparations Using a Coupled Receptor-Binding/RP-HPLC Assay

Selective Capture and Determination of Receptor-Binding Hemagglutinin in Influenza Vaccine Preparations Using a Coupled Receptor-Binding/RP-HPLC Assay

The expanding role of mass spectrometry in the field of vaccine development

Selective reversed-phase high-performance liquid chromatography method for the determination of intact SARS-CoV-2 spike protein

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