• Title/Summary/Keyword: Single radial immunodiffusion

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Immunogenic characterization of AlPO4 adsorbed Td vaccine and liposome-mediated Td vaccine

  • Remees Shuhsadhe;Junise Vazhayil;Heyam Saad Ali;Hiba Orsud;Ahmed Elmontaser Omer Mergani
    • Clinical and Experimental Vaccine Research
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    • v.12 no.3
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    • pp.232-239
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    • 2023
  • Purpose: The purpose of this study was to compare the antigenic potency and stability of tetanus and diphtheria (Td) vaccines when combined with aluminum phosphate (AlPO4) and liposome adjuvants. Materials and Methods: In vitro and in vivo analyses were conducted using the single radial immunodiffusion method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Td vaccines were prepared with AlPO4 adsorption and liposome-mediated delivery, and protein antigens were characterized using these methods. Results: The results revealed that the liposome-mediated Td vaccines exhibited higher immunogenicity compared to the AlPO4-adsorbed Td vaccines. Additionally, the liposome-mediated Td vaccines demonstrated higher stability as native antigens. Conclusion: This study highlights the importance of utilizing liposome adjuvants in vaccine development. The liposome-mediated Td vaccines showed enhanced immunogenicity and stability, making them a promising approach for improving vaccine efficacy. Understanding and optimizing adjuvant strategies can contribute to the development of effective vaccines against various diseases.

Purification and Properties of Osteopontin from Bovine Milk (우유로부터 Osteopontin의 분리.정제 및 특성에 관한 연구)

  • Choi, K.W.;Kim, D.W.;Lee, S.W.
    • Journal of Animal Science and Technology
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    • v.45 no.3
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    • pp.491-498
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    • 2003
  • The purpose of this study is to observe purification and properties of osteopontin(OPN) from bovine milk. The purification of osteopontin from bovine milk was performed by using ion-exchange and hydrophobic chromatography. SDS-PAGE analysis revealed that the protein migrated at Mw. 60,000. NH2-terminal sequence analysis of the first seven amio acids revealed the protein to be identical to that previously reported for bovine OPN. 35-wk-old chickens, including 3 Single Comb White Leghorn (SCWL), were used to produce egg yolk antibody(IgY) against OPNas a antigen. However, the anti-OPN antibody activities determined by ELISA. Immunological assy of OPN in milk was performed using radial immunodiffusion test based on the standard curve of pure OPN. The radial precipitation lines of four different milk samples indicated that the concentrations of OPN in the milk samples were within the range of 31.7 to 39.7${\mu}g$/ml. On inhibition with OPN on precipitation of calcium phosphate, OPN was slightly higher than casein phosphopeptide(CPP) and poly-glutamic acid.

A STUDY OF SALIVARY IMMUNOGLOBULIN AND DENTAL CARIES (타액 면역글로부린과 치아우식의 상관관계에 관한연구)

  • Kim, Jung-Sik
    • Restorative Dentistry and Endodontics
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    • v.9 no.1
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    • pp.37-49
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    • 1983
  • This study was undertaken to observe the relationship between salivary IgA, IgG, agglutinin titer & dental caries. The subjects divided into two groups, an active caries group (AC group) and caries-free and treated caries group (CFTC group). The AC group consisted of 36 subjects who had one or more carious lesions and the CFTC group of 12 subjects who had no evidence of caries or had filled teeth without present carious teeth for the last six months. The IgA, IgG and IgM levels in their saliva were measured by single radial immunodiffusion method using a disposable low-level immunodiffusion plate. The salivary agglutinin titers to Streptococcus mutans were measured by microtitration system. The results were as follow; 1. The mean value of IgA concentration in saliva of AC group was slightly higher than that of CFTC group, but its difference was slight. 2. The mean value of IgG concentration in saliva of AC group was slightly lower than that of CFTC group. The IgM concentration in saliva of both groups was neither below 1.1 mg/dl nor detected on LC partigen immunodiffusion plate. 3. There was no difference in the agglutinin titer to S. mutans antigen by serotypes, but low level agglutinin to type d was measured in both groups. 4. AC group showed low correlation between IgA, IgG & DMFT, but CFTC group revealed negative correlation. 5. The relationship between salivary IgA & agglutinin titers to S. mutans was low correlation in AC group, but CFTC group showed significant positive correlation. (P<0.05) 6. There were no specific correlations among the concentrations of salivary immunoglobulins, agglutinin titers to S. mutans, and the DMF teeth. They had no close concern to induce the dental caries.

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Salivary Immunoglobulin A Concentrations in Patients with Oral Ulcer (구강궤양 환자의 타액내 면역 글로블린 A의 농도)

  • Geun-Chun Ryu;Chang-Lyuk Yoon
    • Journal of Oral Medicine and Pain
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    • v.19 no.1
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    • pp.117-124
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    • 1994
  • Parotid and whole saliva were collected from 27 healthy adults, from 25 years of age to 30, and from 27 patients with oral ulcer, from 23 years of age to 61. The amount of each Salivary immunoglobulin A was measured by single radial immunodiffusion (SRID) technique. Results were as follows : 1. There was no significant difference between the normal group and the disease group in the concentration of immunoglobulin A in whole saliva. 2. The concentration of immunoglobulin A in parotid saliva of the normal group was higher than the disease group and the difference was statistically significant between the two groups. (p<0.01) 3. The concentration of immunoglobulin A of the parotid saliva in both groups was higher than that of the whole saliva.

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A Study on pH, Viscosity, Microorganisms and Immunoglobulin A of the Saliva (타액의 수소이온 농도, 점도, 세균 및 면역글로블린 A에 관한 연구)

  • Jin-Woo Sohn;Sung-Woo Lee
    • Journal of Oral Medicine and Pain
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    • v.17 no.2
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    • pp.87-97
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    • 1992
  • The purpose of this study is to investigate the age-and sex-related changes in the pH of resting saliva, viscosity, microorganisms and immunoglobulin A of stimulated whole saliva, and to investigate their correlations. The 120 healthy subjects were included in this study and the author used cone-and plate digital viscometer for viscosity, MSB agar for Streptococcus mutans, SL Rogosa agar for lactobacilli, and single radial immunodiffusion technique for immunoglobulinA. The obtained results were as follows : 1. There was no significant difference in pH, viscosity, Streptococcus mutans lactobacilli and immunoglobulin A of the saliva between males and females. 2. The viscosity values of stimulated whole saliva showed the increasing pattern with aging. 3. DMFS (or dmfs) rate was not correlated with pH, viscosity, Streptococcus mutans, lactobacilli and immunoglobulin A of the saliva. 4. There was a significant difference in the concentration of immunoglobulin A between the group under 10 and groups above 10. 5. The viscosity values of stimulated whole saliva showed the increasing pattern with decreasing of the number of Streptococcus mutans.

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Evaluation of Haemagglutinin Content by RP-HPLC to Generate Pandemic Influenza Vaccine

  • Kang, Hyunkyung;Roh, Hang Sik;Song, Hyemin;Lee, Kwangmoon;Chung, Seung-Tae;Ban, Sang-ja;Mo, In Pil;An, Beum-Soo;Ahn, Chi-Young
    • Toxicological Research
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    • v.32 no.4
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    • pp.269-274
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    • 2016
  • The potency of influenza vaccine is determined based on its hemagglutinin (HA) content. In general, single radial immunodiffusion (SRID) assay has been utilized as the standard method to measure HA content. However, preparation of reagents for SRID such as antigen and antibody takes approximately 2~3 months, which causes delays in the development of influenza vaccine. Therefore, quantification of HA content by other alternative methods is required. In this study, we measured HA contents of H1N1 antigen and H1N1 influenza vaccine by reverse phase-high performance liquid chromatography (RP-HPLC) methods. The presence of HA1 and HA2 was investigated by silver staining and Western blot assay. In addition, accuracy and repeatability of HA measurement by RP-HPLC were evaluated. Comparison of HA concentration by SRID and RP-HPLC revealed a precise correlation between the two methods. Our results suggest that RP-HPLC assay can replace SRID in the event of a pandemic flu outbreak for rapid vaccine development.

The Production of Egg Yolk Immurnoglobulin (IgY) Raised against 3T3L-1 Cell Membrane Protein and the Control of Adipocytes Differentiation (3T3L-1세포의 막단백질에 대한 난황면역글로뷸린 (IgY)의 생산과 지방세포의 분화조절작용)

  • 김상윤;황성구;구의섭;고태송
    • Korean Journal of Poultry Science
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    • v.26 no.3
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    • pp.179-188
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    • 1999
  • The present was undertaken to establish a model for the control of adipocytes differentiation by using antibody from egg yolk. The emulsion of membrane protein of 3T3L-1 cell membrane protein with the complete Freund's adjuvant was firstly immunized in layer. Second and third boosting were undertaken with two weeks intervals by injection of the emulsion of the same antigen with the incomplete Freund's adjuvant. After 4 week of the first immunization, eggs were collected and antibody (IgY) was purified from egg yolk. The purity of IgY was 60-98% determined by single radial immunodiffusion (SRID) methods. Titer value of the antibody showed high reactiviy for the preadipocytes membrane protein measured by ELISA. When the IgY was added in the test media containing either 2.5% porcine serum or 10% FBS(control), the differentiation of 3T3L-1 cells and Glycerol-3-phosphate dehydrogenase(GPDH) activities was significantly decreased compared to the control cells(p〈0.05). When mice were subcutaneously injected with IgY raised against membrane protein of 3T3L-1 cells for 3 weeks, adipose tissue mass around ovary was tended to be decreased in female mice compared to those of control mice. It is suggested that a potential for manipulating of lipid accumulation through decrease in 3T3L-1 cell differentiation and fat accumulation in female mice by IgY treatment.

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계배 발생에 따른 닭 alpha-Fetoprotein의 분석

  • 박대규;유정아
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.314-321
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    • 1989
  • 배의 발생과 분화에 따른 혈청 alpha-fetoprotein (AFP)의 생리적 기능 규명을 위한 기초실험의 일환으로, 모체와는 완전히 독립된 상태에서 발생과 분화가 이루어지는 계배의 혈청을 재료로 하여 CM-Sephadex C-50, hydroxyapatite 및 DEAE-Sepharose A-25등의 크로마토그라피 방법에 의해 닭 혈청 AFP를 분리하였다. 계 AFP는 분자량이 약 60 Kd로 사람 및 다른 포유동물 AFP의 분자량 (64-74 Kd)보다 작았다. 닭 AFP, 계배혈청 및 닭 혈청 등을 토끼에 면역하여 항체를 제조하였고, 이를 이용하여 닭의 배발생 단계에 따른 혈청 AFP 함량의 변화를 전기영동 및 면역학적 방법으로 정성 및 정량 분석을 실시하였다. 혈청 AFP 함량은 발생 7일-배 (AFP 농도 : 0.81 mg/ml)부터 점차 증가하여 13일-배에서 최고치인 2.46 mgfml로 나타났으며 그 이후 급격히 감소되어 부화 직후에는 매우 낮은 농도 (0.22mg/ml)였고 부화 4일에 거의 없어 졌으며, 이와 같은 AFP 함량의 변화는 혈청 알부민 함량의 변화와 거의 반비례 함을 알았다. For the preliminary step to investigate the site of AFP synthesis, the ontogenic characteristics and the physiological function of AFP, alpha-fetoprotein was isolated from chick embryo serum through the procedures of CM-Sephadex C-50, hvdroxvapatite and DEAE-Sephadex A-50 chro-matography. The isolated AFP was proved to be pure and its molecular weight was found to be about 60 Kd. With this purified AFP, rabbit anti-chick hor was produced. The serum AFP level in chick embryo has been investagated by using polyacrylamide gel electrophoresis, immunoelec-trophoresis and single radial immunodiffusion from 7 days of incubation until 4 days after hatch-ing. It was found that the AFP level is increased from 0.81 mg/ml at daw-7 embryo to maximum value of 2.46 mg/ml at day-13 embryo, followed by rapid decreases until hatching.

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Comparison of diagnostic methods on failure of passive immunoglobulin transfer to Korean-indigenous calves (한우 송아지의 초유 섭취 수준 측정 진단법 비교)

  • Lee, Byoung-Seok;Kang, Mun-Il;Chung, Yong-Un;Lee, Chai-Yong;Han, Dong-Un;Wee, Sung-Hwan;Yoon, So-Rah;Cho, Jae-Jin;Kang, Ju-Won
    • Korean Journal of Veterinary Service
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    • v.31 no.4
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    • pp.505-519
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    • 2008
  • For screening the appropriate field diagnostic techniques to failure of passive immunoglobulin transfer(FPT) in Korean-indigenous calves, 258 sera was examined by spectinophotometry for total protein(TP) and globulin(Glo), sodium sulfate precipitation test(SSPT), zinc sulfate turbidity test(ZSTT), and single radial immunodiffusion test(sRID). All calves aged within 6-week old. Morbidity and mortality to various diseases, mainly including enteric and respiratory disorders, were 18.9%(49) and 4.2%(11), respectively. FPT was 27,9%(72/258) when the cutoff point of TP was $4.5g/d{\ell}$ and among them the morbidity and mortality were 27.9% and 6.9%, respectively. FPT was 29.1%(75/258) when the cutoff point of Glo was $2.0g/d{\ell}$ and among them the morbidity and mortality were 29.0% and 6.9%, respectively. FPT was 13.1%(34/258) when the cutoff point of SSPT was 1+ and among them the morbidity and mortality were 67.6% and 23.5%, respectively. FPT was 19.7%(51/258) when the cutoff point of IgG with sRID was $1,000mg/d{\ell}$ and among them the morbidity and mortality were 41.1% and 11.7%, respectively. In addition, mean concentration of IgG with sRID tested was $2,150mg/d{\ell}$ at 3-day old but $1,100mg/d{\ell}$ at 9-days with $1,100mg/d{\ell}$. The results of the study were suggested that SSPT for FPT was the relatively reliable and convinient method for evaluating the immune status of calves(P<0.05).