Multiplexed lateral flow assay for detection of HPV associated cancer
AssigneeArizona Board of Regents on behalf of Arizona State University
InventorsKaren Anderson, Ching-Wen Hou
Life of this patent
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Abstract
Methods and compositions are provided for the detection of HPV infection in a subject sample. In particular, the methods and composition include variants of certain HPV proteins (e.g., HPV16 E6 protein), wherein the HPV proteins have been modified to enhance solubility while maintaining or enhancing seropositivity to HPV positive plasma/sera. Additional methods and compositions relate to improving purification of HPV proteins through the use of modified amino acid sequences. These approaches may enhance the performance of serological detection techniques and support broader implementation of detection assays.
Claims15
A composition comprising (a), (b), and (c), linked to a solid support:
- (a) human papillomavirus type 16 (HPV16) E6 variant polypeptides, wherein the variant polypeptides consist of:
- (i) a variant of SEQ ID NO:
- 1 having a deletion of amino acids 30-35 of SEQ ID NO:
- 1 (HPV16 E6 (Del30-35)),
- (ii) a fragment of SEQ ID NO:
- 1 consisting of amino acids 70-100 of SEQ ID NO:
- 1 HPV16 E6 (70-100)),
- (iii) a fragment of SEQ ID NO:
- 1 consisting of amino acids 87-158 of SEQ ID NO:
- 1 (HPV16 E6 (87-158)), and
- (iv) a fragment of SEQ ID NO:
- 1 consisting of amino acids 1-86 of SEQ ID NO:
- 1 ((HPV16 E6 (1-86)),
- (b) HPV16 E2 polypeptide consisting of SEQ ID NO:
- 4, and
- (c) HPV E7 polypeptide (SEQ ID NO:
- 2).
A nucleic acid molecule, wherein the nucleic acid molecule comprises one or more sequences encoding human papillomavirus type 16 (HPV16) E6 or E2 variant polypeptides, wherein the variant polypeptides consist of:
- (a) a variant of SEQ ID NO:
- 1 having a deletion of amino acids 30-35 (HPV16 E6 (Del30-35)) of SEQ ID NO:
- 1;
- (b) a fragment of SEQ ID NO:
- 1 consisting of amino acids 70-100 of SEQ ID NO:
- 1 (HPV16 E6 (70-100));
- (c) a fragment of SEQ ID NO:
- 1 consisting of amino acids 87-158 of SEQ ID NO:
- 1 (HPV16 E6 (87-158));
- (d) a fragment of SEQ ID NO:
- 1 consisting of amino acids 1-86 of SEQ ID NO:
- 1 ((HPV16 E6 (1-86));
- (e) SEQ ID NO:
- 4.
A method for detecting HPV antibodies, the method comprising:
- (a) contacting an antibody-containing sample from a subject to (i), (ii), and (iii):
- (i) human papillomavirus type 16 (HPV16) E6 variant polypeptides, wherein the variant polypeptides consist of:
- (A) a variant of SEQ ID NO:
- 1 having a deletion of amino acids 30-35 (HPV16 E6 (Del30-35)) of SEQ ID NO:
- 1,
- (B) a fragment of SEQ ID NO:
- 1 consisting of amino acids 70-100 of SEQ ID NO:
- 1 (HPV16 E6 (70-100)),
- (C) a fragment of SEQ ID NO:
- 1 consisting of amino acids 87-158 of SEQ ID NO:
- 1 (HPV16 E6 (87-158)), and
- (D) a fragment of SEQ ID NO:
- 1 consisting of amino acids 1-86 of SEQ ID NO:
- 1 ((HPV16 E6 (1-86)):
- (ii) HPV16 E2 polypeptide consisting of SEQ ID NO:
- 4;
- and
- (iii) HPV E7 polypeptide (SEQ ID NO:
- 2);
- to form a binding reaction;
- (b) incubating the binding reaction under conditions to allow antibody to bind the HPV16 polypeptides and form a polypeptide-antibody complex;
- (c) detecting the complex.
Dependencies are inferred from the claim text.