Lateral Flow Immunoassay for Qualitative Detection of Insulin Antibodies

Anthony Jerez

Anthony's first successful bands!

Anthony's first successful bands!

Undergraduate Student Project

Introduction

Many point of care test strips like pregnancy test strips are available over the counter. Diabetic patients that are taking medications such as insulin can sometimes develop a tolerance.  Nevertheless, the test strip I am developing will allow diabetics to determine what is occurring inside their body and what their next step should be!

Abstract

Administration of insulin for diabetic patients has been a long standing effective treatment that overtime may lose efficacy. There have been cases in which insulin treatment losses effectiveness due to an immune response trigger of insulin antibody production. For doctors to determine if this is the reason insulin treatment has stopped working, lab tests need to be conducted to determine insulin antibody presence and concentration. Lateral flow immunoassay (LFIA) has been effective for point of care (POC) testing with cost efficient and quick results. The insulin antibodies fab region binds to the antigen, insulin, which is conjugated to the gold nanoparticle (AuNPs) via a EDC-NHS reaction. Now binded to the AuNPs, it carries the analyte, insulin antibodies, to two immobilization sites the test line (TL) and the control line (CL). The insulin antibodies have the second fab region available to bind to the immobilized insulin on the testline. Furthermore, rabbit anti-insulin polyclonal antibody is used for the control line. Currently, qualitative detection of insulin antibodies has been achieved, but clear immobilization of the TL and CL need to be optimized. Overall, quantitative studies need to be performed to determine the sensitivity of the assay.

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