Aav vector and assay for anti-aav (adeno-associated virus) neutralizing antibodies
AssigneeThe Children's Hospital of Philadelphia
InventorsKatherine A. HIGH, Federico MINGOZZI, Yifeng CHEN
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Abstract
Virus vectors, virus particles, and methods and uses of screening for, detecting, analyzing and determining amounts of virus antibody, or neutralizing antibody activity of samples are provided. Such virus vectors, virus particles, and methods and uses are applicable to a broad range of virus types, such as lentiviruses, adenovirus, and adeno-associated virus (AAV) serotypes. Methods and uses include virus antibody screening, such as anti-virus immunoglobulins screened for, detected, analyzed and amounts determined.
Claims70
A recombinant AAV vector sequence, wherein the vector sequence comprises a reporter transgene, wherein the reporter transgene comprises (a) a single-stranded or a self-complementary genome, (b) is operably linked to one or more expression regulatory elements, and (c) is flanked by one or more flanking elements.
A recombinant AAV vector, wherein the vector comprises a reporter transgene, wherein the reporter transgene comprises (a) a single-stranded or a self-complementary genome, (b) is operably linked to one or more expression regulatory elements, and (c) is flanked by one or more flanking elements, wherein the recombinant AAV vector comprises an infectious recombinant AAV particle.
A method for analyzing for or detecting or measuring antibodies that bind to AAV, comprising:
- (a) providing infectious recombinant AAV particles that encapsidate a recombinant vector, wherein (i) the vector comprises a reporter transgene, (ii) the reporter transgene comprises a single-stranded or a self-complementary genome, and (iii) the reporter transgene is operably linked to one or more expression regulatory elements and flanked by one or more flanking elements;
- (b) providing a biological sample from a subject for analyzing or detecting antibodies that bind to AAV;
- (c) providing cells that can be infected with said infectious recombinant AAV particles;
- (d) contacting or incubating the infectious recombinant AAV particles of (a) with the biological sample of (b) thereby producing a resulting mixture (M);
- (e) contacting the cells of (c) with resulting mixture (M) under conditions in which the infectious recombinant AAV particles of (a) can infect and express the reporter transgene in said cells;
- (f) measuring expression of the reporter transgene;
- and
- (g) comparing said reporter transgene expression of (f) to reporter transgene of a control, said control either a negative (−) control (i) lacking antibodies that bind to AAV, or (ii) having a predetermined amount of antibodies that bind to AAV, thereby analyzing for or detecting or measuring antibodies that bind to AAV.
The AAV vector or method of claim 27 or 28, wherein the mutated, modified or variant AAV ITR has a deleted D sequence, and/or a mutated, modified or variant terminal resolution site (TRS) sequence.
Dependencies are inferred from the claim text.