Article Title: Validation of Serological Antibody Profiles Against Human Papillomavirus Type 16 Antigens as Markers for Early Detection of Cervical Cancer
Figure Lengend Snippet: Seropositivity to E4, E7, and VLPs-L1 in serum of healthy women and women with CC by Slot blot system. In vitro purified HPV antigens were immobilized in PROTRAN membranes using the Hybri-Slot system, and membranes were fixed and blocked as described in “Materials and Methods.” Strips containing the 3 HPV16 antigens (E4 10 ng, E7 10 ng, VLPs-L1 200 ng) and negative controls (buffer alone and GFP 200 ng) were incubated with dilutions (1:2,500) of sera from women with CC (A), sera from healthy women (B), and positive and negative female serum controls for anti-HPV antibodies (C). Biotinylated secondary antibody (dil 1:10,000) and Streptavidin-horseradish peroxidase (1:8,000) were used to develop the system, and the bands were finally visualized by chemiluminescence in the Odyssey Fc system. CC = cervical cancer, GFP = green fluorescent protein, HPV = human papillomavirus, VLPs-L1 = virus-like particles from L1 protein.
Article Snippet: The biotinylated secondary antibody (goat anti-human immunoglobulins; dilution 1:10,000) (Jackson Immunoresearch Laboratories, USA) was diluted in blocking solution and incubated at 4o C for 1.5 hours.
Techniques: Dot Blot, In Vitro, Purification, Incubation