• 제목/요약/키워드: BSA (Albumin, from bovine serum)

검색결과 152건 처리시간 0.032초

판토텐산의 분석을 위한 효소면역측정법 (An Enzyme-Linked Immunosorbent Assay for Detection of Pantothenic Acid)

  • 손동화;박윤식;배근원
    • 한국식품과학회지
    • /
    • 제32권5호
    • /
    • pp.1009-1014
    • /
    • 2000
  • PA를 분석하기 위하여 효소면역측정법을 개발하고자 하였다. Bc방법과 Po방법으로 BSA에 PA를 conjugation하여 각각의 PA-BSA conjugate(PA-BSA[Bc]와 PA-BSA[Po])를 제조하였으며, 이를 토끼에 면역하여 항PA-BSA 항체를 얻었다. 항PA-BSA[Po] 항체를 사용한 ciELISA의 결과에서 경합반응이 제대로 일어나지 않았기 때문에, 식품 속에 있는 PA를 검출하기 위해서 PA-BSA[Po]을 코팅한 후 항PA-BSA[Bc] 항체를 사용하였다. 이 결과에서 PA의 검출한계가 1 ppm인 것을 확인할 수 있었으며, 교차반응을 통해 PA 유도체들에 대해서도 항PA-BSA[Bc] 항체가 PA에 대해 특이성이 매우 강하였다. 또한, MBA의 결과에서는 그 검출한계가 10ppb인 것을 확인할 수 있었다. 분석시료인 계란(109%), 상추(64%), 소간(344%)의 식품시료에 대한 실험에서 상추를 제외하고는 ciELISA는 MBA의 결과와 비교해 볼 때 양호한 결과를 얻을 수 있었다. 그러므로, ciELISA는 MBA보다 분석시간, 교차반응 등의 면에서 장점이 있어 식품 중 PA의 검출에 효과적으로 활용할 수 있을 것이다.

  • PDF

난황 항체의 안정성에 관한 연구 (Studies on the Stability of Hen′s Egg Yolk Immunoglobulins)

  • 이경애
    • 한국식품조리과학회지
    • /
    • 제12권1호
    • /
    • pp.54-59
    • /
    • 1996
  • Immunoglobulins (IgY) were isolated from egg yolk of hens immunized with bovine serum albumin(BSA). The stability of anti-BSA IgY against heat and pH was investigated. Antibody activity was measured by enzyme linked immunosorbent assay. IgY was relatively heat-stable and most of the antibody activity remained after heating up 65$^{\circ}C$ for 30 minutes. IgY was stable at pH 5-11. However, inactivation of IgY was observed below pH 4, or above pH 12. Inactivation of IgY proceeded rapidly at low pHs(pH 2-3). Most of the antigen binding activity was lost at low pHs probably because of some conformational changes.

  • PDF

Candida pseudotropicalis의 원형질체 융합: 원형질체 형성 및 재생과 융합 조건 (Protoplast fusion of Candia Pseudotropicalis: The conditions for protoplast formation, regeneration and fusion)

  • 전순배;정기철;배석
    • 미생물학회지
    • /
    • 제24권3호
    • /
    • pp.243-250
    • /
    • 1986
  • Candida pseudotropicalis CBS 607에서의 야생형과 영양요구성 돌연변이 균주에 대한 원형질체 형성 및 재생과 상보적 영양요구성 균주간의 융합에 관해서 연구하였다. 야생형과 his tidine 혹은 adenine요구성 균주에서 원형질체 형성율은 96-100%이였으나 methionine과 tryptophan요구성 균주는 각각 52%와 72%였다. Myoinositol (0.5 mg/ml)을 첨가한 배지에서 전배양한 methionine과 tryptophan조 구성 균주는 원형질체 형성시 bovine serum albumin (BSA, 4 mg/ml)을 첨가하면 96-98%의 원형질체 형성율이 얻어졌다. 원형질체 재생 빈도는 18-20% 였으나. BSA와 myoinositol을 첨가하면 adenine요구성 균주(약 20%)을 제외한 나머지 영양요구성 균주에서는 재생율이 20% 이상 증가했다. 원형질체 융합에 대한 PEG와 $CaCl_2$,최적농도는 20%와 100mM이었고 최적 pH와 반 응시간은 각각 6.0과 30분이 있다. 상보적 영양요구성 균수의 융합빈도는 $1.5{\times}10^{-3}\;to\;8.8{\times}10^{-3}$ 였으며 원행점체의 재생 증진으로 인해 융합율이 현저히 증가했다. Histidine요구성 균주와 tryptophan 요구성 균주을 융합시켰을 때 몇개의 융합세포를 얻었는데 측정된 DNA양과 액상의 차이로 보아 aneuploid나 diploid상태임을 알 수 있었다.

  • PDF

Chlorogenic Acid의 면역보조제 효과 (Immunoadjuvant Activity of Chlorogenic Acid)

  • 한용문
    • 약학회지
    • /
    • 제54권6호
    • /
    • pp.494-499
    • /
    • 2010
  • We have been focussing on discovery of natural compounds that have immunoregulatory activities for many years. In the present study, we investigated if chlorogenic acid (CRA), a polyphenolic compound, has an immunoadjuvant activity. Prior to examining the immunoadjuvant activity, effect of CRA on proliferation of T- or B-lymphocyte was determined. Results showed that CRA enhanced the proliferation of those lymphocytes in dose-dependant manner (P<0.05), and the proliferation enhancement by CRA was appeared to be more effective to B-cells than to T-cells. Based on these observations, it was tested with bovine serum albumin (BSA) and Candida albicans cell wall (CACW) as antigenic sources if CRA has an immunoadjuvant activity. In experiments, BSA alone or a mixture of BSA plus CRA was injected intraperitoneally to mice (BALB/c strain). For a negative control, mice were given only diluent (DPBS) by the same route. In other experiment, CACW was tested by the same way as did with BSA. Three weeks after the first immunization these animals were boosted. Antisera collected from the mice one week after the booster were analyzed by ELISA. Results displayed that the induction of anti-BSA antibody was increased in mice that received the mixture of BSA and CRA as compared to anti-BSA induction in BSA only-given mice groups (P<0.05). In case of CACW, a similar observation as did with BSA was made, resulting in that there was app. 40% increased production of the anti-CACW antiserum from the combination (CACW plus CRA)-received mice as compared to antiserum induction from CACW alone-given animals. Taken all together, these data indicate that CRA has an ability of enhancing antibody production regardless of nature of antigenic sources. Presumably, activation of B-cell proliferation by CRA may plays an important role in the immunoadjuvant activity of the polyphenolic compound.

Addition of Macromolecules to PZM-3 Culture Medium on the Development and Hatching of In vitro Porcine Embryos

  • Kim, J.Y.;Kim, S.B.;Park, M.C.;Park, H.;Park, Y.S.;Park, H.D.;Lee, J.H.;Kim, J.M.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제20권12호
    • /
    • pp.1820-1826
    • /
    • 2007
  • In this study, we conducted various experiments in order to develop enhanced cultural conditions for in vitro-produced porcine embryos. All embryos were produced by in vitro maturation (IVM) and fertilization (IVF) of immature oocytes from abattoir-derived ovaries. In experiment 1, we cultured IVF embryos in 4 different groups, namely, 0% bovine serum albumin (BSA), 3% BSA, 0.05% Polyvinyl alcohol (PVA), and 0.5% Polyvinylpyrrolidone (PVP) added to the basal fluid cultural medium, Porcine zygote medium 3 (PZM-3). The rates of embryo development were higher in the group where the PZM-3 media had been supplemented with 3% BSA than the other groups. While not statistically significant, the percent of blastocysts and hatched blastocytes were 6.9% and 25.0% in the 3% BSA group vs. 1.2-6.4% and 0-16.7% in the other groups, respectively. In experiment 2, we added 10% fetal bovine serum (FBS) to PZM-3 on day 0 of culture and observed the development rate of blastocysts per day of culture from days 0 to 5. The development rate of blastocysts was higher at 15.6% on day 4 than on any other day, and was significantly higher than on day 0 or day 1 (p<0.05). The development rate of hatched blastocysts was 26.7% on day 4, and was higher than on any other day. In experiment 3, we cultured IVF embryos with different fluid culture media, grouped as 1) PZM-3+0.3% BSA (day0-day7); 2) PZM-3+0.3% BSA${\rightarrow}$day-4) PZM-3+10% FBS; 3) PZM-3+0.3% BSA${\rightarrow}$PZM-3+0.3% BSA+(day-4) FBS 10%; and 4) PZM-3+0.3% BSA+10% FBS (day0-day7). The development rates of blastocysts and hatched blastocysts were 21.5% and 53.1% in group 3, respectively, which was significantly higher than group 4 with respect to blastocyst development (5.2%, p<0.05) but not hatched blastocysts (14.3%). The total cell number (TCN) of blastocysts in group 3 was higher at $37.8{\pm}16.1$ than the other groups at $16.8{\pm}4.4$ - $30.1{\pm}10.9$; however, this was not significantly different. The results of this study showed that PZM-3 containing 0.3% BSA and supplemented with FBS during the later stage of culture on day 4 resulted in better TCNs and an increased rate of hatched blastocysts.

Roles of Fucoidan, an Anionic Sulfated Polysaccharide on BSA-Stabilized Oil-in-Water Emulsion

  • Kim, Do-Yeong;Shin, Weon-Sun
    • Macromolecular Research
    • /
    • 제17권2호
    • /
    • pp.128-132
    • /
    • 2009
  • Fucoidan, a sulfated polysaccharide derived from brown seaweed, is an important material valued for its various biological functions, including anti-coagulation, anti-aging, and immune system support. In this study, we examined the potential of fucoidan as a novel emulsifying agent in BSA (bovine serum albumin)-stabilized emulsion at a neutral pH. We measured the dispersed oil-droplet size, surface zeta-potential and creaming formation of 0.5 wt% BSA emulsion (20 wt% oil traction) in the absence and presence of fucoidan. The average particle size and zeta-potential value were 625.4 nm and -30.91 mV in only BSA-stabilized emulsion and 745.2 nm and -44.2 mV in 1.0 wt% fucoidan-added BSA emulsion, respectively. This result suggested that some positive charges of the BSA molecules interacted with the negative charges of fucoidan to inhibit the flocculation among the oil droplets. The creaming rate calculated from the backscattering data measured by Turbiscan dramatically decreased in 1.0 wt% fucoidan-added BSA emulsion during storage. Accordingly, the repulsion forces induced among the oil particles coated with 1.0 wt% fucoidan in emulsion solution resulted in significantly increased emulsion stability. The turbidity of the BSA-stabilized emulsion at 500 nm decreased during five days of storage. However, the fucoidan-added BSA emulsion exhibited a higher value of turbidity than the BSA-stabilized emulsion did. In conclusion, an anionic sulfated fucoidan lowered the surface zeta-potential of BSA-coated oil droplets via the electrostatic interaction, and subsequently inhibited the flocculation among the oil droplets, thereby clearly minimizing the creaming and phase separation of the emulsion.

Immunochemical Studies on Expression of Quinoproteins in Escherichia coli

  • Ryou, Chong-Suk;Kim, Jae-Beom;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
    • /
    • 제10권1호
    • /
    • pp.95-98
    • /
    • 2000
  • An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

  • PDF

Retention Characteristics of Various Proteins in Flow Field-Flow Fractionation : Effects of pH, Ionic Strength, and Denaturation

  • 송종희;김원숙;박영훈;유의경;이대운
    • Bulletin of the Korean Chemical Society
    • /
    • 제20권10호
    • /
    • pp.1159-1164
    • /
    • 1999
  • The study investigated the effect of carrier composition (ionic strength and pH) on the retention of various proteins in flow field-flow fractionation (Flow FFF) as well as the conformational change of Bovine Serum Albumin (BSA) with urea concentration, storage time and temperature. The study found that the retention of protein in Flow FFF increased with the ionic strength of the carrier liquid. Most proteins were well solubilized at pH = 7-8. The hydrodynamic diameters obtained from Flow FFF retention data agree well with theoretical values. The retention increased and the peak shape became distorted at extreme pH conditions of the carrier solution. The selected carrier composition for comparison between the literature value of proteins was 0.05 M tris buffer solution with a pH of 8. Storing BSA at 4 ±2℃ over a period of three months resulted in slow dimerization. Also, in case of the storage of BSA at 37 ±5℃ for one week, the retention of both BSA monomer and dimer increased with the urea concentration. Finally, the structural composition of specific enzymes: malonyl-CoA decarboxylase (MCDC) and malonyl-CoA synthesis (MCS) was determined by using Flow FFF at specific carrier solutions. The molecular weight of the natural MCDC was determined to be 208 kDa, which means it is a homotetramer, while that of the MCS was determined to be 47 kDa, which means it is a monomer.

기니픽과 마우스에서 신규 퀴놀론 항균제 DW-116의 항원성 시험 (Antigenicity of DW-116, a New Quinolone Antibiotic, in Guinea Pigs rind Mice)

  • 권현진;한형미;이흠숙;정용호;윤성호;이문선;이덕근
    • Biomolecules & Therapeutics
    • /
    • 제6권2호
    • /
    • pp.165-170
    • /
    • 1998
  • Antigenic potential of DW-116, a newly synthesized fluoroquinolone, was examined by conduc-ting active systemic anaphylaxis (ASA), passive cutaneous anaphylaxis (PCA) and passive hemagglutination (PHA) tests. In ASA test, mild to moderate signs of anaphylactic responses were observed in the groups sensitized with low (2 mg/body) and high (10 mg/body) doses of DW-116 alone and the group sensitized with DW-116 plus adjuvant. Some moderate to severe anaphylactic reactions were observed in the group sensitized with a DW-116-bovine serum albumin (BSA) conjugate plus adjuvant when challenged with a DW-116-guinea pig senHn albumin (GSA) conjugate. However these reactions were considered to be a cross-reaction between BSA and GSA since similar reactions were induced when challenged by GSA alone. In heterologous PCA test using mice and rats, positive responses were not detected in any of the experimental groups. In PHA test, positive responses were observed in the groups sensitized with low and high doses of DW-116 alone and the group sensitized with DW-116 plus adjuvant. However, these responses were not considered to be drug-specific because some positive responses were also seen in the negative control group. From these results, it was concluded that DW-116 is not likely to have specific antigenic potential in clinical use.

  • PDF

술폰화 PP-g-Styrene 중공사 이온교환막의 합성과 BSA 단백질 분리에 관한 연구 (Synthesis of Sulfonated Hollow PP-g-Styrene Fibrous Ion-exchange Membrane and Separation of BSA Protein)

  • 황택성;이진혁
    • 폴리머
    • /
    • 제26권4호
    • /
    • pp.415-421
    • /
    • 2002
  • E-beam 전조사법을 이용하여 HPP-g-styrene 공중합체와 술폰화 반응을 통한 술폰화 HPP-g-styrene 섬유이온교환체를 합성하였다. 그라프트율은 스티렌 단량체 농도가 증가함에 따라 증가하였으며 스티렌 단량체 농도가 80%에서 그라프트율이 128%로 최대를 나타냈다. 술폰화율은 그라프트율이 증가함에 따라 증가하는 경향을 나타내었으며, 그라프트율이 100%일 때 13.4%로 최대값을 나타내었다. 술폰화 HPP-g-styrene 섬유이온교환체의 이온교환용량은 약 3.42 meq/g으로써 흡착 성능이 매우 우수한 소재임을 확인하였다. BET 분석결과 술폰화 HPP-g-styrene의 비표면적은 62.54 $m^2/g$, 기공크기는 25 $\AA$으로 반응전보다 비표면적은 감소하였고 기공크기는 약간 증가하는 경향을 보였다. 또한 Bovine Serum Albumin (BSA) 흡착 실험 결과 술폰화도가 증가함에 따라 BSA 흡착 용량이 증가하는 경향을 나타내었으며, 술폰화도 13.4%에서 BSA 흡착용량 3.8 mg/g으로 최대를 나타내었다. 따라서 본 연구에서 합성한 섬유이온교환체가 BSA 흡착.분리에 적합한 소재임을 확인하였다.