Bioconjugation of AuNPs with HPV 16/18 E6 antibody through physical adsorption technique

Abstract

Gold nanoparticle (AuNP) bioconjugates are increasingly being utilised in biomedicine due to their low toxicity on biological tissues and unique electronic and chemical properties. They have been utilised in several biological applications namely manufacture of nanomaterials, biosensing, electron microscopy and drug delivery systems. Particularly, immuno-assays often employ gold nanoparticles (AuNPs) to enhance detection of a biological component. This paper presents a study on the bioconjugation of AuNPs with an antibody against an oncoprotein that is overexpressed in cervical carcinoma progression through physical adsorption. The study successfully developed a HPV 16 E6/18 E6-HRP (CP15)-AuNPs bioconjugate. The study also demonstrated that the antibody pI, gold colloidal solution pH and amount of antibody determine the generation of stable Antibody-AuNPs bioconjugates. These results further demonstrate that physical adsorption can be used to produce functional Antibody-AuNPs that could be employed in diagnostic immunoassays.

Development of HPV 16/18 E6 oncoprotein paper-based nanokit for enhanced detection of HPV 16/18 E6 oncoprotein in cervical cancer screening

“…T1085 Beijing Solarbio Science & Technology Co. Ltd) and in a 10 cm Petri dish for 1 hour at room temperature. The membranes were incubated with HPV 16 E6/18 E6-HRP (CP15)-AuNPs bioconjugate (25) to allow formation of an immune complex at a concentration of 4 μg/ml for 30 min at room temperature. The paper strips were washed three times each about 5 minutes with TBS-T and incubated with TMB reagent (Thermofisher).…”

“…The nanokit uses the direct ELISA-based immunoassay principle as illustrated in Figure 2. oncoprotein analyte using the dot blot molecular technique (25). A dot blot is a technique similar to western blot as they both utilize antibodies to identify specific membrane bound proteins.…”

“…Gold nanoparticle (AuNP) bioconjugates are increasingly being utilised in biomedicine due to their low toxicity on biological tissues and unique electronic and chemical properties. In a recent study, the bioconjugation of AuNPs with HPV 16 E6/18 E6-HRP (CP15) antibody against HPV 16/18 E6 oncoprotein that is overexpressed in cervical carcinoma progression was developed through physical adsorption (25). This study therefore, developed a paper-based nanokit through evaluation of the immune sensing mechanism and diagnostic accuracy of the HPV 16 E6/18 E6-HRP (CP15)-AuNPs bioconjugate in enhanced detection of E6 oncoprotein in cervical cancer screening.…”

“…Gold nanoparticle (AuNP) bioconjugates are increasingly being utilised in biomedicine due to their low toxicity on biological tissues and unique electronic and chemical properties. In a recent study, the bioconjugation of AuNPs with HPV 16 E6/18 E6-HRP (CP15) antibody against HPV 16/18 E6 oncoprotein that is overexpressed in cervical carcinoma progression was developed through physical adsorption (25).…”

Untitled

“…Gold nanoparticle (AuNP) bioconjugates are increasingly being utilised in biomedicine due to their low toxicity on biological tissues and unique electronic and chemical properties. In a recent study, the bioconjugation of AuNPs with HPV 16 E6/18 E6-HRP (CP15) antibody against HPV 16/18 E6 oncoprotein that is overexpressed in cervical carcinoma progression was developed through physical adsorption [25]. Hydrogen peroxidase is a colorimetric enzyme, used in the detection of analytes through a color change observed via a naked eye, when the substrate 3,3',5,5'-Tetramethylbenzidine (TMB) is added to the reaction [26].…”