• 제목/요약/키워드: preliminary purification

검색결과 36건 처리시간 0.025초

吸着 크로마토그라피法에 의한 사람 ALpha-Fetoprotein의 分離 (Affinity Purification of Human Alpha-Fetoprotein)

  • Kang, Shin-Sung;Bang, Ok-Sun;Park, Tae-Kyu
    • 한국동물학회지
    • /
    • 제29권4호
    • /
    • pp.283-293
    • /
    • 1986
  • 사람의 alpha-fetoprotein(AFP)에 대한 모노클론 항체의 생산 및 분석을 위하여, 태아조직을 재료로 추출법, DEAE-cellulose 및 concanavalin A-Sepharose, Cibacron blue F3GA-agarose, immunoadsorbent column등의 흡착크로마토그라피법에 의해 AFP를 분리하였다. 총 534g의 태아조직에서 AFP의 量은 8.76 mg으로서 순수분리되었음을 확인하였다.

  • PDF

Human Chorionic Gonadotropin(hCG)에 대한 단일콜론항체 생산 : 단일클론항체의 분리정제 및 그 특성조사와 hCG정량을 위한 Immunochemiluminometric assay(ICMA)개발 (Production of Monoclonal Antibody to Human Chorionic Gonadotropin(hCG) : Purification and Properties of a Monoclonal Antibody, and Immunochemiluminometric assay(ICMA) for the Assay of hCG)

  • 최상훈;이병철;오재욱;이용환;서광영;정길생;김종배
    • 한국가축번식학회지
    • /
    • 제12권1호
    • /
    • pp.51-62
    • /
    • 1988
  • hCG로 면역화된 생쥐의 비장세포와 골수종양세포(SP 2/0 Ag14)를 융합하여 hCG에 대한 단일클론항체를 생산하는 잡종세포(hybridoma)를 얻었다. 생산된 면역글로부린 type과 titer 그리고 면역분석법 이용시 감도 등을 조사함으로써 그 특성을 조사하였다. 배지나 복수내에 존재하는 항체를 순수분리 정제하기 위하여 gel-filtration, DEAE-ion exchange chromatography, 그리고 affinity chromatography 원리에 의한 방법 등을 사용하여 전기영동(SDS-PAGE)으로 순수도를 조사함으로 상호 비교하였다. 또한 hCG의 정량을 위하여 항체를 plastic tube에 피복시킨 것과 화학발광체로 표지된 항체를 이용한 소위 two-site immunochemiluminometric assay(ICMA)를 개발하여 생산된 항체의 이용가능성을 제시하였다.

  • PDF

일라이트와 이산화티탄을 활용한 콘크리트 블록의 수질환경개선을 위한 실험연구 (A Study on Experiment for Improvement of Water Quality in Concrete Block Using Illite and TiO2 Powder)

  • 허재원;강경수;김영수
    • 한국구조물진단유지관리공학회 논문집
    • /
    • 제22권1호
    • /
    • pp.115-122
    • /
    • 2018
  • "일라이트와 이산화티탄을 활용한 콘크리트 블록의 수질환경개선을 위한 실험연구"를 진행하기 위해 모르타르 예비실험, 수질정화특성, 방오실험, 어독성 실험 및 실 콘크리트 블록 제조 후 관련 KS에 준한 실험을 진행한 결과 다음과 같은 결론을 얻었다. KS에 준한 콘크리트 블록의 성능평가 결과 전 조건에서 시험기준치의 압축강도를 상회하는 것으로 측정되었으며, 다공질구조의 일라이트 치환에 따른 흡수율 증가가 문제점으로 예상되었으나, 이산화티탄과 사전 혼합하여 치환함으로써 일라이트의 대형 공극내 이산화티탄이 정착함에 따라 흡수율 또한 문제가 없는 것으로 관찰되었다.

장내세균의 시간차 혼합배양이 보여주는 균수측정의 비교 (Colony Count with Mixed Culture of Enteric Bacteria by in vitro Quantitative Method)

  • 황선철;전보성
    • 미생물학회지
    • /
    • 제11권4호
    • /
    • pp.175-180
    • /
    • 1973
  • This study was attempted to see more clear relationships among the enterobacteria, especially between the intestinal normal flora and pathogenic bacteria. It has been known that some intestinal normal flora produce the bactrial metabolites that are harmful to other enteric bacteria. One of the metabolites is known as colicin, the protein fraction, which possesses certain degree of inhibitory effect against other bacterial growth fraction, whih possesses certain degree of inhibitory effect against other bacterial growth. As a preliminary study for a colicin purification, the antagonistic effect of E, coli to groups of Salmonella and Shigella has been studied by means of in vitro quantitative culture method. 1. E.coli showed definite inhibitory effects aganist both Salmonella and Shigella groups in the mixture of two organisms. 2. The inhibitory effects of E.coli in the E.coli-Salmonella and the E.coli-Shigella mixture occurred from 4 hours incubation following the inoculation. 3. Even the complete inhibition of pathogenic enteric bacterial growth was noticed in the E.coli-Salmonella mixture at overnight incubation. 4. Among the diluted mixtures, 1:100, 1:1,000, and 1:10,000, survival rate of pathogenic enteric bacteria in the mixtures with E.coli showed least affected at the 1:1,000 dilution. 5. It was found that the antagonistic effect aganist groups of Salmonella-shigella was depending upon the groups of the genera.

  • PDF

Overexpression, Purification, and Preliminary X-ray Crystallographic Analysis of Human Brain-Type Creatine Kinase

  • Bong, Seung-Min;Moon, Jin-Ho;Jang, Eun-Hyuk;Lee, Ki-Seog;Chi, Young-Min
    • Journal of Microbiology and Biotechnology
    • /
    • 제18권2호
    • /
    • pp.295-298
    • /
    • 2008
  • Creatine kinase (CK; E.C. 2.7.3.2) is an important enzyme that catalyzes the reversible transfer of a phosphoryl group from ATP to creatine in energy homeostasis. The brain-type cytosolic isoform of creatine kinase (BB-CK), which is found mainly in the brain and retina, is a key enzyme in brain energy metabolism, because high-energy phosphates are transfered through the creatine kinase/phosphocreatine shuttle system. The recombinant human BB-CK protein was overexpressed as a soluble form in Escherichia coli and crystallized at $22^{\circ}C$ using PEG 4000 as a precipitant. Native X-ray diffraction data were collected to $2.2{\AA}$ resolution using synchrotron radiation. The crystals belonged to the tetragonal space group $P4_32_12$, with cell parameters of a=b=97.963, $c=164.312{\AA},\;and\;{\alpha}={\beta}={\gamma}=90^{\circ}$. The asymmetric unit contained two molecules of CK, giving a crystal volume per protein mass $(V_m)$ of $1.80{\AA}^3\;Da^{-1}$ and a solvent content of 31.6%.

Structural Characterization of Growth-Related Translationally Controlled Tumor Protein P23

  • Lee, Bong-Jin;Hong, Yoon-Hun;Park, Sang-Ho;Lee, Kyunglim
    • 한국자기공명학회논문지
    • /
    • 제5권1호
    • /
    • pp.46-55
    • /
    • 2001
  • P23, a translationally controlled turner protein is involved in the interleukin-4 secretion from human basophils and is also known to be an IgE-dependent histamine-releasing factor. However, the precise physiological function and structure of P23 have not been elucidated. In the current study, we constructed the optimal expression and purification protocol of P23 and investigated the secondary structure and structural stability in various conditions. Circular dichroism (CD) investigation showed that the secondary structure of P23 adopts mainly a P-sheet conformation. CD spectroscopy and differential scanning calorimetry revealed that P23 is fairly stable in the pH range of neutral and mild-basic conditions and in the temperature range of 10 - 50$\^{C}$. Since the thermal stability and the P-sheet content of P23 were decreased by the addition of Ca$\^$2+/ ion, it could be suggested that Ca$\^$2+/ion induces structural change by partially destabilizing the structure of P23. In addition various H experiments were monitored to solve the aggregation of P23. Den results will provide the preliminary structural information about P23.

  • PDF

치자(梔子)(Gardenia Fructus)를 이용(利用)한 식용색소(食用色素) 개발(開發)에 관(關)한 연구(硏究) (A Study on the Development of Food Dye from Gardenia Fructus)

  • 서화중
    • Journal of Nutrition and Health
    • /
    • 제14권1호
    • /
    • pp.26-33
    • /
    • 1981
  • Crocin was extracted from Gardenia fructus in order to manufacture a natural food dye. In the extraction and purification process methanol and ethanol were used as the most Suitable solvent selected in preliminary test and 9.28%g/100g of Cretin product in 99.89% of purity was acquired. The lethal dose of Cretin administered in mouse by intra peridoneal was 5.36g/kg which is much lower toxic than any other. The toxic dose which caused diarrhoea in rat was 2.55g/kg and maximum no effect level was found to be 272mg/kg. From the $1%{\sim}2.5%$ content of Crocin in rat diet reduction of body weight appeared and GTP and GOT (Transaminase) value in creased significantly. Crocin showed a good properties of tolerating acid, alkali, sunlight, reductant, oxidant, and salt compared with tar dyes. The range from 100 ppm to 500ppm bring out bright $yellow{\sim}orange$ color, the most effective color, with a good solubility in water-slightly in oil. The acceptable daily intake of crocin was calculated as 2.72mg/kg based on maximum no effect dose (2% additive dose).

  • PDF

Overexpression, Purification, and Preliminary X-Ray Crystallographic Studies of Methionine Sulfoxide Reductase B from Bacillus subtilis

  • Park, Ae-Kyung;Shin, Youn-Jae;Moon, Jin-Ho;Kim, Young-Kwan;Hwang, Kwang-Yeon;Chi, Young-Min
    • Journal of Microbiology and Biotechnology
    • /
    • 제18권1호
    • /
    • pp.59-62
    • /
    • 2008
  • The peptide methionine sulfoxide reductases (Msrs) are enzymes that catalyze the reduction of methionine sulfoxide back to methionine. Because of two enantiomers of methionine sulfoxide (S and R forms), this reduction reaction is carried out by two structurally unrelated classes of enzymes, MsrA (E.C. 1.8.4.11) and MsrB (E.C. 1.8.4.12). Whereas MsrA has been well characterized structurally and functionally, little information on MsrB is available. The recombinant MsrB from Bacillus subtilis has been purified and crystallized by the hanging-drop vapor-diffusion method, and the functional and structural features of MsrB have been elucidated. The crystals belong to the trigonal space group P3, with unit-cell parameters a=b=136.096, $c=61.918{\AA}$, and diffracted to $2.5{\AA}$ resolution using a synchrotron-radiation source at Pohang Light Source. The asymmetric unit contains six subunits of MsrB with a crystal volume per protein mass $(V_M)\;of\;3.37{\AA}^3\;Da^{-1}$ and a solvent content of 63.5%.

Crystallization and Preliminary X-Ray Crystallographic Analysis of PAS Factor from Vibrio vulnificus

  • Lee, Jun Hyuck;Kim, Soo Young;Rho, Seong-Hwan;Im, Young Jun;Kim, Young Ran;Kim, Mun-Kyoung;Kang, Gil Bu;Rhee, Joon Haeng;Eom, Soo Hyun
    • Molecules and Cells
    • /
    • 제20권3호
    • /
    • pp.361-363
    • /
    • 2005
  • Plasmid Achromobacter secretion (PAS) factor is a putative secretion factor that induces the secretion of periplasmic proteins. PAS factor from Vibrio vulnificus was crystallized at 294 K by the hanging drop vapor-diffusion method. It was isolated as a monomer during the purification procedures. The native crystal belongs to the F222 space group with unit cell parameters a = 56.1, b = 74.4, $c=80.0{\AA}$, ${\alpha}={\beta}={\gamma}=90^{\circ}$. The crystal was soaked in cryoprotectant containing 1 M NaBr for 1 h for MAD phasing. The diffraction limit of the Br-MAD data set was $1.9{\AA}$ using synchrotron X-ray irradiation at beam line BL-18B at the Photon Factory, Japan.

Characterization of a Blend-Biosurfactant of Glycolipid and Lipopeptide Produced by Bacillus subtilis TU2 Isolated from Underground Oil-Extraction Wastewater

  • Cheng, Fangyu;Tang, Cheng;Yang, Huan;Yu, Huimin;Chen, Yu;Shen, Zhongyao
    • Journal of Microbiology and Biotechnology
    • /
    • 제23권3호
    • /
    • pp.390-396
    • /
    • 2013
  • Biosurfactants have versatile properties and potential industrial applications. A new producer, B. subtilis TU2, was isolated from the underground oil-extraction wastewater of Shengli Oilfield, China. Preliminary flask culture showed that the titer of biosurfactant obtained from the broth of TU2 was ~1.5 g/l at 48 h (718 mg/l after purification), with a reduced surface tension of 32.5 mN/m. The critical micelle concentration was measured as 50 mg/l and the surface tension maintained stability in solution with 50 g/l NaCl and 16 g/l $CaCl_2$ after 5 days of incubation at $70^{\circ}C$. FT-IR spectra exhibited the structure information of both glycolipid and lipopeptide. MALDI-TOF-MS analyses confirmed that the biosurfactant produced by B. subtilis TU2 was a blend of glycolipid and lipopeptide, including rhamnolipid, surfactin, and fengycin. The blended biosurfactant showed 86% of oil-washing efficiency and fine emulsification activity on crude oil, suggesting its potential application in enhanced oil recovery.