Glycidyl methacrylate-co-N-vinyl-2-pyrrolidone coated polypropylene strips: synthesis, characterization and standardization for dot-enzyme linked immunosorbent assay.
Glycidyl methacrylate and N-vinyl-2-pyrrolidone (GMA-co-NVP) copolymers with various GMA:NVP ratios were synthesized by solution polymerization technique in toluene using 2,2'-azobisisobutyronitrile (AIBN) as free radical initiator and dip coated onto polypropylene strips. The copolymer composition in polymeric coatings was confirmed by proton NMR spectroscopy. Various techniques like FTIR, SEM and contact angle were used for surface characterization of the polymer coatings. These polymer coated strips were evaluated and standardized for their application in dot-ELISA in two steps. In first step, specificity, sensitivity and reproducibility of the assay on developed polymer coated strips was evaluated through a model system using rabbit anti-goat IgG, goat anti-rabbit IgG and goat anti-rabbit IgG HRP (horseradish peroxidase)-conjugate. Polymer coating with GMA-NVP mol% ratio of 78:22 was able to detect rabbit anti-goat IgG antibody at a concentration as low as 2 ng mL(-1) with 1% BSA as blocking agent using antispecies IgG peroxidase conjugate diluted 1500 times. In the second step, the sensitivity and specificity of the developed system was established with human blood and finally used to identify the source of mosquito blood meal which is an important parameter in epidemiological studies, particularly in determining the role of mosquito in malaria transmission. The time duration of standardized assay with developed polymer coated strips was cut down to one hour compared to the 3-4h required in usual dot-ELISA.
Tyagi
Charu
C
Centre for Biomedical Engineering, Indian Institute of Technology, Delhi, India. tyagicharu11@rediffmail.com
Tomar
Lomas
L
Singh
Harpal
H
D000818:Animals; D000906:Antibodies / Q000097:blood; D009033:Culicidae / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards*; D005502:Food; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008855:Microscopy, Electron, Scanning; D011126:Polypropylenes / Q000737:chemistry*; D011205:Povidone / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D011485:Protein Binding / Q000187:drug effects; D011934:Reagent Strips / Q000737:chemistry*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012995:Solubility; D017550:Spectroscopy, Fourier Transform Infrared
D000906:Antibodies; D011126:Polypropylenes; D011934:Reagent Strips; C534929:glycidyl methacrylate-co-N-vinyl-2-pyrrolidone; D011205:Povidone