Identification of peptide sequences as a measure of Anthrax vaccine stability during storage
نویسندگان
چکیده
The UK anthrax vaccine is an alum precipitate of a sterile filtrate of Bacillus anthracis Sterne culture (AVP). An increase in shelf life of AVP from 3 to 5 years prompted us to investigate the in vivo potency and the antigen content of 12 batches with a shelf life of 6.4 to 9.9 years and one bulk with a shelf life of 23.8 years. All batches, except for a 9.4-year-old batch, passed the potency test. Mass spectrometry (MS) and in-gel difference 2-dimensional gel electrophoresis (DIGE) were used to examine antigens of the pellet and supernatant of AVP. The pellet contained proteins with a MW in excess of 15 kDa. DIGE of desorbed proteins from the pellet revealed that with aging, 19 spots showed a significant change in size or intensity, a sign of protein degradation. MS identified 21 proteins including protective antigen (PA), enolase, lethal factor (LF), nucleoside diphosphate kinase, edema factor, and S-layer proteins. Fifteen proteins were detected for the first time including metabolic enzymes, iron binding proteins, and manganese dependent superoxide dismutase (MnSOD). The supernatant contained131 peptide sequences. Peptides representing septum formation inhibitor protein and repeat domain protein were most abundant. Five proteins were shared with the pellet: 2,3,4,5-tetrahydropyridine-6-dicarboxylate N-succinyltransferase, enolase, LF, MnSOD, and PA. The number of peptide sequences increased with age. Peptides from PA and LF appeared once batches exceeded their shelf life by 2 and 4 years, respectively. In conclusion, changes in antigen content resulting from decay or desorption only had a limited effect on in vivo potency of AVP. The presence of PA and LF peptides in the supernatant can inform on the age and stability of AVP.
منابع مشابه
Stability and potency studies of anthrax vaccine (Bacillus anthracis 34F2 Sterne strain) in Iran
For evaluation the stability and potency of anthrax vaccine prepared in Razi Vaccine and Serum Research Institute in Iran, samples of different batches of vaccine were kept at 4-8 °C (refrigerator), 20-25 °C (room temperature) and 37 °C (incubator).The viable spores/ml of vaccines were determined using plate–counting method before and after holding at different temperatures monthly. Vaccine p...
متن کاملDesign of cocktail peptide vaccine against Cytomegalovirus infection
Objective(s):Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immuno suppressed patients. Therefore, significant effort has been made towards the development of a vaccine. In this study, the expression of the pp65 and gB fusion peptides and Fc domain of mouse IgG2a as a novel delivery system for selective uptake of antigens by antigen-presenting cells (APCs) in Pich...
متن کاملIn silico identification of epitopes from house cat and dog proteins as peptide immunotherapy candidates based on human leukocyte antigen binding affinity
The objective of this descriptive study was to determine Felis domesticus (cat) and Canis familiaris (dog) protein epitopes that bind strongly to selected HLA class II alleles to identify synthetic vaccine candidate epitopes and to identify individuals/populations who are likely to respond to vaccines. FASTA amino acid sequences of experimentally validated allergenic proteins of house cat and d...
متن کاملNovel Delivery Systems for Nasal Administration of Anthrax Vaccine
Sheena Hailin Wang: Novel Delivery Systems for Nasal Administration of Anthrax Vaccine (Under the direction of Dr. Anthony J. Hickey, Ph.D., D.Sc.) There is current biodefense interest in protection against Anthrax. The inhaled form of anthrax is difficult to diagnose and can be fatal without prompt antibiotic intervention. Prophylactic vaccination has become a critical measure for protection o...
متن کامل