Self-assembling recombinant papillomavirus capsid proteins
AssigneesThe United States of America as represented by the Department of Health and Human Services, Deutsches Krebsforschungszentrum, The German Cancer Research Center
InventorsDouglas Lowy, John Schiller, Reinhard Kirnbauer
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Abstract
Recombinant papillomavirus capsid proteins that are capable of self assembly into capsomer structures and viral capsids that comprise conformational antigenic epitopes are provided. The capsomer structures and viral capsids, consisting of the capsid proteins that are expression products of a bovine, monkey or human papillomavirus L1 conformational coding sequence proteins, can be prepared as vaccines to induce a high titer neutralizing antibody response in vertebrate animals. The self assembling capsid proteins can also be used as elements of diagnostic immunoassay procedures for papillomavirus infection.
Claims46
A genetic construct, comprising a papillomavirus L1 conformational coding sequence, inserted into a baculovirus transfer vector, and operatively expressed by a promoter of that vector.
A method for producing a recombinant papillomavirus capsid protein, assembled into a capsomer structure, or a portion thereof, comprising the steps of:
- cloning a papillomavirus gene that codes for an L1 conformational capsid protein into a transfer vector wherein the open reading frame of said gene is under the control of the promoter of said vector;
- transferring the recombinant vector into a host cell, wherein the cloned papillomavirus gene expresses said papillomavirus capsid protein;
- and
- isolating capsomer structures, comprising said papillomavirus capsid protein, from said cell.
A virus capsid or a capsomer structure, or a portion thereof, consisting essentially of papillomavirus L1 capsid protein.
A unit dose of a vaccine, comprising a peptide having conformational epitopes of a papillomavirus L1 capsid protein, or L1 protein and L2 capsid proteins, in an effective immunogenic concentration sufficient to induce a papillomavirus neutralizing antibody titer of at least about 103 when administered according to an immunizing dosage schedule.
A method for immunizing a vertebrate against papillomavirus infection, comprising administering to said vertebrate a recombinant genetic construct comprising a conformational papillomavirus L1 coding sequence, and allowing said coding sequence to be expressed in the cells or tissues or said vertebrate, whereby an effective, neutralizing, immune response to papillomavirus is induced.
A method of detecting humoral immunity to papillomavirus infection in a vertebrate comprising the steps of:
- (a) providing an effective antibody-detecting amount of a papillomavirus capsid peptide having at least one conformational epitope of a papillomavirus capsomer structure;
- (b) contacting the peptide of step (a) with a sample of bodily fluid from a vertebrate to be examined for papillomavirus infection, and allowing papillomavirus antibodies contained in said sample to bind thereto, forming antigen-antibody complexes;
- (c) separating said complexes from unbound substances;
- (d) contacting the complexes of step (c) with a detectably labelled immunoglobulin-binding agent;
- and
- (e) detecting anti-papillomavirus antibodies in said sample by means of the labelled immunoglobulin-binding agent that binds to said complexes.
A method of detecting papillomavirus in a specimen from an animal suspected of being infected with said virus, comprising:
- contacting said specimen with antibodies having a specificity to one or more conformational epitopes of the capsid of said papillomavirus, said antibodies having a detectable signal producing label, or being attached to a detectably labelled reagent;
- allowing said antibodies to bind to said papillomavirus;
- and
- determining the presence of papillomavirus present in said specimen by means of said detectable label.
A papillomavirus infection diagnostic kit, comprising capsomer structures consisting essentially of papillomavirus L1 capsid protein, or capsomer structures comprising papillomavirus L1 protein and L2 capsid proteins, or antibodies to either of said capsomers structures, singly or in combination, together with materials for carrying out an assay for humoral or cellular immunity against papillomavirus, in a unit package container.
Dependencies are inferred from the claim text.