• Title/Summary/Keyword: Radial immunodiffusion

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Detection of antibodies in swine serum to Aujeszky's disease virus using agar-gel immunodiffusion test (Agar-gel immunodiffusion test를 이용한 돼지 혈청중 Aujeszky's disease virus 항체 검출에 관한 연구)

  • Cho, Hyo-gueon;Jun, Moo-hyung
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.297-307
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    • 1990
  • To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

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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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A Systematic Study on Selected Amentiferous Plant Taxa - By Quantitative Analysis of Proteins - (유이화서 식물군의 통계분류학적 연구 - 단백질의 정량분석적 접근 -)

  • 이유성
    • Journal of Plant Biology
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    • v.28 no.3
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    • pp.207-216
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    • 1985
  • Radial immunodiffusion, rocket immunoelectrophoresis, and Boyden procedure have been employed as quantitative analysis of pollen proteins in a systematic investigation of selected amentiferous plant taxa. Data presented here are continued and supplementary to the previous qualitative analysis of immunoprecipitin systems for the same purpose. Although the number of taxa tested has been limited, the serological evidence indicates that the Betulaceae has the greatest similarity to the Fagaceae, next to the Juglandaceae, the least to the Salicaceae, when antisera against Alnus hirsuta and Betula platyphylla var. japonica were used for tests. Within the Betulaceae Alnus and Betula show greatly similar affinities together, but less similar to the rest of genera: Carpinus, Carya and Corylus. When antisera against Quercus aliena, Q. dentata, and Q. glauca were used for tests, the following decreasing order to serological affinities was obtained: Quercus Alnus, Betula Carpinus, Carya, Corylus Juglans, Pterocarya Populus. Overall serological data come closer to supportint the classification systems of Cronquist, Takhtajan, and Hutchinson; but less of Thorne and Bessey. In addition this investigation indicated that pollen, with its high protein content, provided an excellent source of extractable antigens for serosystematic researches.

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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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Further Systematic Studies on Cornus and Relatives by Immunoelectrophoresis (면역학적 전기영동에 의한 층층나무속과 그 근연군의 계통학적 추가연구)

  • Lee, Yoo-Sung
    • Journal of Plant Biology
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    • v.28 no.2
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    • pp.95-104
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    • 1985
  • Additional serological data from radial immunodiffusion and conventional immunoelectrophoresis with combination of presaturation of antibody were obtained for further interpretation of Cornus (and relatives) systematics. Pollen proteins were compared by qualitative and quantitative means. Cornus drummondi and C. stolonifera are very similar to C. florida, and C. amomum and C. recemosa are less similar. Nyssa constitutes the next distinct and most similar family, Nyssaceae to the Cornaceae. The serological affinities indicate that Corokia and Griselinia should not be included within the Cornaceae. Their taxonomic treatment to elevate to the family level is now awaiting until more data are accumulated.

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Pre- and Postoperative Changes of Serum Levels of Slpha 1-proteinase Inhibitor in Patients with Bronchogenic Cancer (폐암 환자에서의 폐엽절제술 전후의 혈청 Alpha 1-Proteinase Inhibitor의 변화)

  • 이재성
    • Journal of Chest Surgery
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    • v.28 no.3
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    • pp.221-227
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    • 1995
  • Malignancy is one of the several exogenous and endogenous factors that increase serum alpha 1-PI. In fact, serum levels of alpha 1-PI were significantly elevated in the patients with the nonresectable bronchogenic cancer. the purpose of this work was to determine if the immediate postoperative change of serum alpha 1-PI level following tumor resection relates to the patient`s postoperative course. Clinical experimental study was carried out to investigate the postoperative changes of serum alpha 1-PI level following operation for 20 cases of bronchogenic cancer and 10 cases of control, nephrectomy patients Alpha 1-PI concentrations in serum was quantitated by use of radial immunodiffusion technique.The results were as follows ; Preoperative serum level of alpha 1-PI was significantly elevated in patients with bronchogenic cancers [p < 0.001 , when compared to normal control levels. Immediate postoperative serum alpha 1-PI level was significantly increased in patients with bronchogenic cancer [p < 0.05 , but slightly decreased at control groups. The peak serum level of alpha 1-PI was the postoperative three days, and then gradually decreased at the 5, 9, 14 days, but slightly elevated comparing to preoperative alpha 1-PI levels. Serum alpha 1-PI level in patients with adenocarcinoma was elevated, when compared to squamous cell carcinoma, but not significantly. According to the stages of the bronchogenic cancer, each levels of the serum alpha 1-PI were slightly different, but the whole postoperative changes were the general similarity. There were no significant difference in changes of the serum alpha 1-PI level, according to the operative procedures. As the alpha 1-PI is acute reactant, that it was required at the reoperative state of the bronchogenic cancer and rapid response, consumption or requirement were occurred, postoperatively. Therefore, alpha 1-PI can be perioperative indicator for the evaluation of the bronchogenic cancer.

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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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