Production of hybridomas secreting antibodies to the cornea

Zam, Z. Suzanne; Jones, Pauline; Das, Noveen D.

doi:10.3109/02713688109001819

Cited by 5 publications

(2 citation statements)

References 6 publications

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“…Murine monoclonal antibodies. HPV L1-specific mouse mAbs were produced using hybridoma technology 54,55 . BALB/c mice were primely immunized subcutaneously with HPV VLPs formulated with Freund's complete adjuvant (50 μg/ dose) at week 0 and then two boost immunizations using Freund's incomplete adjuvant were implemented at weeks 2 and 4.…”

Section: Methodsmentioning

confidence: 99%

Rational design of a multi-valent human papillomavirus vaccine by capsomere-hybrid co-assembly of virus-like particles

et al. 2020

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The capsid of human papillomavirus (HPV) spontaneously arranges into a T = 7 icosahedral particle with 72 L1 pentameric capsomeres associating via disulfide bonds between Cys175 and Cys428. Here, we design a capsomere-hybrid virus-like particle (chVLP) to accommodate multiple types of L1 pentamers by the reciprocal assembly of single C175A and C428A L1 mutants, either of which alone encumbers L1 pentamer particle self-assembly. We show that co-assembly between any pair of C175A and C428A mutants across at least nine HPV genotypes occurs at a preferred equal molar stoichiometry, irrespective of the type or number of L1 sequences. A nine-valent chVLP vaccine-formed through the structural clustering of HPV epitopes-confers neutralization titers that are comparable with that of Gardasil 9 and elicits minor cross-neutralizing antibodies against some heterologous HPV types. These findings may pave the way for a new vaccine design that targets multiple pathogenic variants or cancer cells bearing diverse neoantigens.

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Section: Methodsmentioning

confidence: 99%

Rational design of a multi-valent human papillomavirus vaccine by capsomere-hybrid co-assembly of virus-like particles

et al. 2020

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“…HPV L1-specific mouse mAbs were produced using the hybridoma technology previously described 49 , 50 . BALB/c mice were immunized subcutaneously three times at 2-week intervals (0, 2, 4 weeks) with nine HPV monovalent vaccines, respectively.…”

Section: Methodsmentioning

confidence: 99%

N-terminal truncations on L1 proteins of human papillomaviruses promote their soluble expression in Escherichia coli and self-assembly in vitro

Wei

Wang

et al. 2018

Emerging Microbes & Infections

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Human papillomavirus (HPV) is the causative agent in genital warts and nearly all cervical, anogenital, and oropharyngeal cancers. Nine HPV types (6, 11, 16, 18, 31, 33, 45, 52, and 58) are associated with about 90% of cervical cancers and 90% of genital warts. HPV neutralization by vaccine-elicited neutralizing antibodies can block viral infection and prevent HPV-associated diseases. However, there is only one commercially available HPV vaccine, Gardasil 9, produced from Saccharomyces cerevisiae that covers all nine types, raising the need for microbial production of broad-spectrum HPV vaccines. Here, we investigated whether N-terminal truncations of the major HPV capsid proteins L1, improve their soluble expression in Escherichia coli. We found that N-terminal truncations promoted the soluble expression of HPV 33 (truncated by 10 amino acids [aa]), 52 (15 aa), and 58 (10 aa). The resultant HPV L1 proteins were purified in pentamer form and extensively characterized with biochemical, biophysical, and immunochemical methods. The pentamers self-assembled into virus-like particles (VLPs) in vitro, and 3D cryo-EM reconstructions revealed that all formed T = 7 icosahedral particles having 50–60-nm diameters. Moreover, we formulated a nine-valent HPV vaccine candidate with aluminum adjuvant and L1 VLPs from four genotypes used in this study and five from previous work. Immunogenicity assays in mice and non-human primates indicated that this HPV nine-valent vaccine candidate elicits neutralizing antibody titers comparable to those induced by Gardasil 9. Our study provides a method for producing a nine-valent HPV vaccine in E. coli and may inform strategies for the soluble expression of other vaccine candidates.

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Stop codon mutagenesis for homogenous expression of human papillomavirus L1 protein in Escherichia coli

Wang

Fan

et al. 2017

Protein Expression and Purification

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Production of hybridomas secreting antibodies to the cornea

Cited by 5 publications

References 6 publications

Rational design of a multi-valent human papillomavirus vaccine by capsomere-hybrid co-assembly of virus-like particles

Rational design of a multi-valent human papillomavirus vaccine by capsomere-hybrid co-assembly of virus-like particles

N-terminal truncations on L1 proteins of human papillomaviruses promote their soluble expression in Escherichia coli and self-assembly in vitro

Stop codon mutagenesis for homogenous expression of human papillomavirus L1 protein in Escherichia coli

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