Development of a biosensor-based method for detection and isotyping of antibody responses to adenoviral-based gene therapy vectors.
A biosensor-based assay, using a surface plasmon resonance detection system, was developed to detect and isotype anti-adenoviral antibodies in patients dosed with an adenoviral-based gene therapy vector. In the assay, whole, intact virus was immobilized onto the sensor chip surface. Electron microscopy and monoclonal antibody studies provide evidence that the virus remains intact after immobilization. The patients tested had preexisting serum levels of anti-adenoviral antibodies. A classic anamnestic response was observed in patients dosed with the gene-therapy agent. Isotyping experiments indicated that IgG antibodies predominated in serum even at the predose time point. Analysis of ascites fluid samples from some patients indicated detectable levels of IgA in addition to IgG. Results obtained using the biosensor-based assay corresponded to an existing enzyme-linked immunosorbent assay. The assay was easy to perform and the automated instrument reduced the required "hands on" time. In addition to studying the development of anti-adenoviral antibodies, the techniques described may be applied to virus:receptor interaction studies or antiviral drug:virus interaction studies.
Abad
Leslie W
LW
Schering-Plough Research Institute, Department of Biotechnology Development, 1011 Morris Avenue, Union, NJ 07083, USA.
Neumann
Meredith
M
Tobias
Lori
L
Obenauer-Kutner
Linda
L
Jacobs
Sheila
S
Cullen
Constance
C
D000256:Adenoviridae / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000914:Antibodies, Viral / Q000032:analysis* / Q000096:biosynthesis / Q000145:classification; D001202:Ascitic Fluid / Q000276:immunology; D015374:Biosensing Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis* / Q000096:biosynthesis / Q000145:classification; D008854:Microscopy, Electron; D020349:Surface Plasmon Resonance / Q000379:methods; D013499:Surface Properties; D013552:Swine; D013997:Time Factors
D000906:Antibodies; D000914:Antibodies, Viral; D007136:Immunoglobulins