An ELISA method to estimate the mono ADP-ribosyltransferase activities: e.g in pertussis toxin and vaccines

• Published in: Analytical biochemistry 2018-01, Vol.540-541, p.15-19
• Main Authors: Asokanathan, Catpagavalli, Tierney, Sharon, Ball, Christina R., Buckle, George, Day, Ami, Tanley, Simon, Bristow, Adrian, Markey, Kevin, Xing, Dorothy, Yuen, Chun-Ting
• Format: Article
• Language: English
• Subjects: 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulphonic acid); ABTS; Adenosine diphosphate; ADP; ADP Ribose Transferases - analysis; ADP Ribose Transferases - antagonists & inhibitors; ADP Ribose Transferases - metabolism; ADP-Ribosyltransferase enzymatic unit; bovine serum albumin; BSA; Chromatography, High Pressure Liquid; Clostridium - enzymology; diphtheria toxoid; dithiothreitol; DTT; ELISA; Enzyme Assays - methods; Enzyme-ELISA; Enzyme-Linked Immunosorbent Assay; Escherichia coli - enzymology; Escherichia coli - metabolism; FHA; filamentous haemagglutinin; fimbriae type 2 and type 3; Fims2/3; formaldehyde; H2O2; Haemophilus influenzae type b; Hib; high performance liquid chromatography; His; HIST; histamine sensitisation test; histidine; horseradish peroxidase; HPLC; HRP; Humans; hydrogen peroxide; inactivated poliovirus vaccine; IPV; LPC; lysophosphatidylcholine; mAb; Macro domain; Mono ADP-Ribosyltransferase; Monoclonal antibody; NAAD; NAD; nicotinamide adenine dinucleotide; nicotinic acid adenine dinucleotide; optical density; OVA; ovalbumin; PBS; Peptides - chemistry; Peptides - metabolism; pertactin; Pertussis toxin; Pertussis Toxin - analysis; Pertussis Toxin - metabolism; Pertussis toxoid; Pertussis vaccines; phosphate buffered saline; PRN; PTd; PTx; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; SHD; single human dose; Substrate Specificity; TBS; tetanus toxoid; Tris buffer saline; Vaccines, Conjugate - analysis; Vaccines, Conjugate - metabolism
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2017.10.025

ADP-ribosyltransferase activities have been observed in many prokaryotic and eukaryotic species and viruses and are involved in many cellular processes, including cell signalling, DNA repair, gene regulation and apoptosis. In a number of bacterial toxins, mono ADP-ribosyltransferase is the main cause of host cell cytotoxicity. Several approaches have been used to analyse this biological system from measuring its enzyme products to its functions. By using a mono ADP-ribose binding protein we have now developed an ELISA method to estimate native pertussis toxin mono ADP-ribosyltransferase activity and its residual activities in pertussis vaccines as an example. This new approach is easy to perform and adaptable in most laboratories. In theory, this assay system is also very versatile and could measure the enzyme activity in other bacteria such as Cholera, Clostridium, E. coli, Diphtheria, Pertussis, Pseudomonas, Salmonella and Staphylococcus by just switching to their respective peptide substrates. Furthermore, this mono ADP-ribose binding protein could also be used for staining mono ADP-ribosyl products resolved on gels or membranes. •ADP-ribosyltransferase activities in biological systems and macro domains.•ELISA to estimate mono ADP-ribosyltransferase in pertussis toxin and its vaccines.•A new assay method that could potentially be used for measuring ADP-ribosyltransferase activities in bacteria.