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Pertussis vaccine purification

Zenglan Li, Yan Zhang, Qi Wang, Zhengjun Li, Yongdong Liu, Songping Zhang, Guifeng Zhang, Guanghui Ma, Jian Luo, Zhiguo Su
Development of acellular pertussis vaccine (aPV) requires purification of several components from Bordetella pertussis. While the components pertussis toxin (PT) and filamentous hemagglutinin (FHA) have been successfully purified, the third component, pertactin, proves to be a difficult target due to its very low concentration. In order to solve its purification problem, we performed the surface potential analysis with GRASP2 program. The results demonstrated that there are two major charge patches, one negative and one positive, which are located separately on this linear protein...
July 25, 2016: Vaccine
Vanessa Zavatti, Hector Budman, Raymond Legge, Melih Tamer
Fluorescence spectroscopy in combination with multivariate statistical methods was employed as a tool for monitoring the manufacturing process of pertactin (PRN), one of the virulence factors of Bordetella pertussis utilized in whopping cough vaccines. Fluorophores such as amino acids and co-enzymes were detected throughout the process. The fluorescence data collected at different stages of the fermentation and purification process were treated employing principal component analysis (PCA). Through PCA, it was feasible to identify sources of variability in PRN production...
June 2016: Bioprocess and Biosystems Engineering
An-Guo Liu, Hua-Zhang Li, Xing-Ke Yan, Tian-You He, Li-Li Kan, Jun-Yan Wang, Li-Li Dong
OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Sanyin" acupoints (bilateral "Jiayin" and "Zhongyin") on the cellular immune function of rats with chronic abacterial prostatitis (CAP), so as to explore its mechanism underlying relieving CAP. METHODS: Forty male Wistar rats were randomly divided into four groups: control, model, medication (Cernilton) and EA (n = 10 rats/group). The CAP model was made by injection of allogeneic prostatein purification liquid + Freund's complete adjuvant (1:1) and diphtheria, pertussis, tetanus (DPT) vaccine (0...
June 2013: Zhen Ci Yan Jiu, Acupuncture Research
Hossein Asgarian-Omran, Ali Akbar Amirzargar, Mohammad Arjmand, Mohammadreza Eshraghian, Behrooz Nikbin, Saeid Eshraghi, Marzieh Mahdavi, Jalal Khoshnoodi, Mahmood Jeddi-Tehrani, Hodjatallah Rabbani, Fazel Shokri
BACKGROUND: Filamentous hemagglutinin (FHA) is one of the most important immunoprotective antigens of Bordetella pertussis (B. pertussis) and a major component of the acellular pertussis vaccine. In the present study, three overlapping recombinant fragments from the immunodominant region of FHA were produced and their immunogenicity was investigated. METHODS: Three overlapping coding sequences of FHA antigen were amplified from B. pertussis genomic DNA by PCR. Amplified fragments were expressed in Escherichia coli (E...
January 2013: Avicenna Journal of Medical Biotechnology
R Brodzik, S Spitsin, N Pogrebnyak, K Bandurska, C Portocarrero, K Andryszak, H Koprowski, M Golovkin
The current diphtheria-tetanus-pertussis (DTP) pediatric vaccine is produced from the corresponding pathogenic bacteria Corynebacterium diphtheriae, Clostridium tetani and Bordetella pertussis; five injected doses of DTaP (acellular) vaccine are required for every child in the standard US vaccination schedule. Because the vaccine is derived from native live sources, adverse effects are possible and production is complex and costly. To address issues of safety, ease of renewability and expense, we used recombinant technology in an effort to develop a subunit DPT vaccine derived in non-pathogenic plant expression systems...
June 8, 2009: Vaccine
Tie Wu, Jingxiu Bi, Yongdong Huang, Yan Zhang, Lijing Sun, Chunbao Sun, Zhiguo Su
The low recovery of pertussis toxin (PT) and the low resolving efficiency of chromatography, due to the instability of PT in low salt condition, are the main challenges for its purification. We aplied 2 mol/L urea to prevent the aggregation and disassociation of PT during the purification by ion-exchange chromatography (IEC) and gel filtration chromatography (GFC). The effect of urea on the purification of PT was studied by ELISA assay and non-reduced SDS-PAGE. The activity recoveries of PT and filamentous hemagglutinin (FHA) in IEC and GFC, the resolution efficiency in GFC and the purities of PT and FHA were improved obviously by adding 2 mol/L urea in the mobile phase...
July 2008: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
Ptnar Atakan Ablay, Erkan Ozcengiz
Bordetella pertussis fimbria antigens play an important role in the adhesion to the cell surfaces, and induce the formation of protective antibodies in the host. The aim of this study was to purify the fimbria proteins by a rapid and easy method which involved Superdex 75 gel filtration chromatography, and to investigate their immunogenicity. The fimbria proteins purified from the different strains showed single protein bands in 12.5% SDS-PAGE (Sodium dodecyl sulphate-polyacrylamide gel electrophoresis) method...
January 2007: Mikrobiyoloji Bülteni
Mahdi Aminian, Sheila Sivam, Chiang W Lee, Scott A Halperin, Song F Lee
Pertussis toxoid, diphtheria toxoid, and tetanus toxoid are key components of diphtheria-tetanus-acellular pertussis vaccines. The efficacy of the vaccines is well documented, however, the vaccines are expensive partly because the antigens are derived from three different bacteria. In this study, a fusion protein (PDT) composed of the immunoprotective S1 fragment of pertussis toxin, the full-length non-toxic diphtheria toxin, and fragment C of tetanus toxin was constructed via genetic means. The correct fusion was verified by restriction endonuclease analysis and Western immunoblotting...
February 2007: Protein Expression and Purification
R P Chuprinina, I A Alekseeva, N A Ozeretskovskiĭ
Modern data substantiating the expediency of the use of acellular pertussis vaccine were analyzed. Serious postvaccinal complications caused by the action of the corpuscular pertussis component of adsorbed DPT vaccine served as the basis for the development of acellular pertussis vaccine (APV). During the period of 1990-1996 as many as 8 international field trials of the effectiveness of APV were carried out. The results of these trials and studies were evaluated in accordance with the unified programs and criteria...
January 2006: Zhurnal Mikrobiologii, Epidemiologii, i Immunobiologii
John A Bogdan, Wei Yuan, Karen O Long-Rowe, Jawad Sarwar, Eric Allen Brucker, M S Blake
The introduction of acellular pertussis vaccines has greatly enhanced the safety profile of vaccines to prevent whooping cough. Pertussis toxin (Ptx) is one component produced by Bordetella pertussis that is contained in all of these vaccines, either in combination with other known pertussis virulence factors or as the sole pertussis component, combined with tetanus and diphtheria toxoids. A hydrogen peroxide toxoid of Ptx has been shown to be efficacious in preventing pertussis infections in a mass vaccination trial and is presently licensed in the United States and Europe (B...
October 2003: Applied and Environmental Microbiology
Song F Lee, Scott A Halperin, Jennifer B Knight, Aaron Tait
Acellular pertussis vaccines typically consist of antigens isolated from Bordetella pertussis, and pertussis toxin (PT) and filamentous hemagglutinin (FHA) are two prominent components. One of the disadvantages of a multiple-component vaccine is the cost associated with the production of the individual components. In this study, we constructed an in-frame fusion protein consisting of PT fragments (179 amino acids of PT subunit S1 and 180 amino acids of PT subunit S3) and a 456-amino-acid type I domain of FHA...
September 2002: Applied and Environmental Microbiology
A M Smith, C A Guzmán, M J Walker
Bordetella pertussis is the causative agent of whooping cough, a contagious childhood respiratory disease. Increasing public concern over the safety of whole-cell vaccines led to decreased immunisation rates and a subsequent increase in the incidence of the disease. Research into the development of safer, more efficacious, less reactogenic vaccine preparations was concentrated on the production and purification of detoxified B. pertussis virulence factors. These virulence factors include adhesins such as filamentous haemagglutinin, fimbriae and pertactin, which allow B...
May 2001: FEMS Microbiology Reviews
M Sakaguchi, S Inouye
Most of the children who showed systemic immediate-type reactions, including anaphylactic shock, to measles, mumps, rubella, and varicella vaccines had IgE antibodies to gelatin; thus we suspected that the allergic symptoms are caused by gelatin antigen, which is usually included in these live-virus vaccines as a stabilizer. We hypothesized that the anti-gelatin IgE is elicited by immunization with DTaP (diphtheria-tetanus-acellular pertussis) vaccines, which contained a small amount of gelatin as a spillover protein after purification of pertussis toxin...
October 2000: Japanese Journal of Infectious Diseases
S A Plotkin
The reputation of vaccination rests on a 200-year-old history of success against major infectious diseases. That success has led to the doctrine of 'for each disease, a vaccine'. Although some diseases have proved frustrating, this doctrine carries considerable truth. However, when one reviews the vaccines now available it is apparent that most successes have been obtained when the microbe has a bacteremic or viremic phase during which it is susceptible to the action of neutralizing antibodies, and before replication in the particular organ to which it is tropic...
November 1999: Comptes Rendus de L'Académie des Sciences. Série III, Sciences de la Vie
J G Kreeftenberg
In comparison with the current whole cell pertussis vaccine, the new generation of acellular pertussis vaccines opens new opportunities to improve the standardization of the product, because well defined and characterized components are used in these new products.However, different compositions, purification and inactivation methods are used by different manufacturers. Consequently the various acellular pertussis vaccines in the world are difficult to compare in a meaningful manner using simple laboratory tests...
June 1999: Biologicals: Journal of the International Association of Biological Standardization
G C Sheu, Y Y Wo, C H Lu
Pertussis toxin (PT), a typical A-B oligomer exotoxin of Bordetella pertussis, has been demonstrated to be an essential protective antigen for acellular pertussis vaccine against whooping cough. In order to investigate the associated functionality ascribed to its components, we have purified A and B oligomers for the activity study. The A oligomer (S1 subunit) of PT was expressed in E. coli B834 (DE3) harboring expression vector (pET-20b) with the insert of S1 coding region and purified by metal-chelating column...
August 1997: Zhonghua Minguo Wei Sheng Wu Ji Mian Yi Xue za Zhi, Chinese Journal of Microbiology and Immunology
C L Ju, G C Sheu, Y Cheng, C H Lu
Pertussis toxin (PT) is the major protective antigen of acellular pertussis vaccine (aP). We have established an optimal culture condition for the growth of B. pertussis and the production of PT in a laboratory scale fermentor. It was found that when the dissolved oxygen in medium was supplied with pure oxygen instead of air, the yield of PT was dramatically increased (i.e. from 2-3 mg/l using air to 8-10 mg/l using pure oxygen). PT was purified by affinity chromatography using hydroxyapatite and fetuin-sepharose columns...
May 1997: Zhonghua Minguo Wei Sheng Wu Ji Mian Yi Xue za Zhi, Chinese Journal of Microbiology and Immunology
D Hozbor, F G Chirdo, M E Rodriguez, C Valverde, O Yantorno
Bordetella pertussis produces extracytoplasmic adenylate cyclase toxin (AC) which has received considerable attention as a potential vaccine candidate. Great interest from laboratories involved in production, purification and quality control of acellular pertussis vaccine is focused on finding an appropriate technique for rapid and accurate quantitation of AC antigen. In this paper a competitive ELISA is proposed. A polystyrene microplate coated with purified AC was incubated with the sample to be tested plus anti-AC serum...
December 1995: Biologicals: Journal of the International Association of Biological Standardization
J B Milstien, B G Gellin, M Kane, J L di Fabio, A Homma
A recently completed survey of 63 manufacturers of diphtheria-tetanus-pertussis (DTP) vaccine and its components in 42 countries shows that there is potentially a large excess installed capacity for DTP production. However, many manufacturers are not producing to capacity, and demand and supply for this vaccine are not matched in individual countries. About half of all countries producing DTP vaccine and its components do not have fully functional national control systems, and some countries are performing none of the critical functions for an effective control of quality...
March 1996: Vaccine
J P Himanen, M Sarvas, I M Helander
The levels of non-protein impurities at different stages of purification of model vaccine proteins produced by Bacillus subtilis were assessed with special emphasis on peptidoglycan-wall teichoic acid and lipoteichoic acid. Intracytoplasmically produced proteins were purified by disrupting the lysozyme protoplasts using osmotic shock, depositing the inclusion bodies by low-speed centrifugation, and washing them with detergent. By this procedure most of the cell envelope-derived impurities could be removed. The final product contained less than 1% (w/w) of neutral sugars, fatty acids, phosphate, hexosamine, diaminopimelic acid and glycerol...
1993: Vaccine
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