• 제목/요약/키워드: immunoglobulin M(IgM)

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다양한 HPLC Column에서의 IgY(Immunoglobulin Yolk) 분리특성 (Separation Characteristics of IgY (Immunoglobulin Yolk) in Various HPLC Columns)

  • 송성문;김인호
    • Korean Chemical Engineering Research
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    • 제50권4호
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    • pp.659-665
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    • 2012
  • 동물 혈청 중의 IgG (Immunoglobulin G)에 해당되는 난황에 포함된 면역 단백질 IgY (Immunoglobulin Yolk)는 식품 단백질로 장내 면역 물질로 중요하다. IgY를 정제하기 위해 신선란의 노른자에 카리지난이나 아라빅검을 전처리 물질로 사용하였다. 전처리 후 FPLC (Fast Protein Liquid chromatography)의 DEAE (Diethylaminoethyl) Sepharose 칼럼에서 이온교환법에 의해 불순물을 제거하여 IgY를 얻고, GF HPLC (Gel Filtration High Performance Liquid Chromatography)로 IgY의 분자량을 측정하고 표준 IgY와 비교하여 IgY 단백질을 동정하였다. GF HPLC에서 IgY의 다양성을 발견하였고 IgY 단백질 군의 다양성을 IE HPLC (Ion Exchange High Performance Liquid Chromatography)에서 AX, CX, SCX 칼럼을 사용하여 pH, NaCl 농도를 바꾸어 조사하였다. AX를 사용하여 0.5M NaCl, pH=8 조건에서 3개의 IgY 피크를 분리하였고, SCX를 이용했을 때 0.5M NaCl, pH=5 조건에서도 3개의 IgY 피크를 분리할 수 있었다.

Immunoglobulin productivity assay를 이용(利用)한 홍삼투여(紅蔘投與) 실험동물(實驗動物)의 IgG, IgM, IgA 비교(比較) 연구(硏究) (The Comparative Study of IgG, IgM, and IgA in Laboratory Animal Administrated Red-ginseng, Using Immunoglobulin Productivity Assay)

  • 이범준;소형진;김재완;류재환
    • 대한한방내과학회지
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    • 제28권4호
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    • pp.886-895
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    • 2007
  • Objective : The immune system is a complex of systems, all of which work together to clear infection from the body. In Korea, red ginsenghas been one of the herbs most widely used to enhance the immune system for thousand of years. More recently, red ginseng has been reported to have many positive effects on the immune system. The purpose of this study was evaluate the effects of Korean red ginseng and Chinese red ginseng on IgG, IgM, and IgA, using immunoglobulin productivity assay. Methods : Male SD rats were separated into 3 groups. We administered Korean red ginseng (KRG) to one group and Chinese red ginseng (CRG) to another, with normal saline for the Control group consecutively and orally for 3 months. The dose of red ginseng was 500mg per day, as a powder with soluble water. Immunoglobulin levels from spleen cell were estimated by ELISA kit. Results : In immunoglobulin productivity assay (cell), the IgG level of the KRG group significantly increased but there was no significant difference in the IgG of the CRG group. The IgM level of the KRG group significantly increased stimulated with PWM. When it was unstimulated, the level of IgM in KRG and CRG increased together. The IgA level of the KRG group significantly increased when it was stimulated with PWM and unstimulated. Conclusion : According to the above results, oral administration of red ginseng for 3 months is considered useful for immunomodulatory effect, and Korean red ginseng may be superior to Chinese red ginseng in that effect.

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드렁허리(Monopterus albus) 혈청내 lgM유사 면역글로불린의 구조적 특성 (Structural Characterization of an IgM-like Immunoglobulin in the Serum of Swamp Eel, Monopterus albus)

  • 손영종;이정우;장정순
    • 한국동물학회지
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    • 제33권1호
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    • pp.103-107
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    • 1990
  • BSA로 면역시킨 드렁허리의 혈청에서 lgM 유사면역글로불린의 정제한 후 그 구조적 특성을 구명하였다. 면역후 항혈청에서는 항원특이 항체능이 증가되었다. 정제된 lgM 유사면역글로불린은 199kD의 분자량을 갖는 단량체 4개가 결합된 800kD의 단백질이었으며, 단량체는 70kD인 H-사슬 2개와 29.5kD인 L-사슬 2개가 결합된 형태였다. 또한 드렁허리의 lgM 유사면역글로불린은 lgM의 특성과 같이 포유류 적혈구에 대하여 약한 응집력을 나타냈다.

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Plant-derived PAP proteins fused to immunoglobulin A and M Fc domains induce anti-prostate cancer immune response in mice

  • Yang Joo Kang;Deuk-Su Kim;Seyoung Kim;Young-Jin Seo;Kisung Ko
    • BMB Reports
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    • 제56권7호
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    • pp.392-397
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    • 2023
  • In this study, recombinant Fc-fused Prostate acid phosphatase (PAP) proteins were produced in transgenic plants. PAP was fused to immunoglobulin (Ig) A and M Fc domain (PAP-IgA Fc and PAP-IgM Fc), which were tagged to the ER retention sequence KDEL to generate PAP-IgA FcK and PAP-IgM FcK. Agrobacterium-mediated transformation was performed to produce transgenic tobacco plants expressing four recombinant proteins. Genomic PCR and RT-PCR analyses confirmed the transgene insertion and mRNA transcription of PAP-IgA Fc, PAP-IgM Fc, PAP-IgA FcK, and PAP-IgM FcK in tobacco plant leaves. Western blot confirmed the expression of PAP-IgA Fc, PAP-IgM Fc, PAP-IgA FcK, and PAP-IgM FcK proteins. SEC-HPLC and Bio-TEM analyses were performed to confirm the size and shape of the plant-derived recombinant PAP-Fc fusion proteins. In mice experiments, the plant-derived IgA and IgM Fc fused proteins induced production of total IgGs including IgG1 against PAP. This result suggests that IgA and IgM Fc fusion can be applied to produce recombinant PAP proteins as a prostate cancer vaccine in plant expression system.

A Comparative Study Between Cytomegalovirus Immunoglobulin M-Positive and CMV Immunoglobulin M-Negative Biliary Atresia in Infants Attending a Tertiary Care Hospital in Bangladesh

  • Akter, Sharmin;Karim, ASM Bazlul;Mazumder, Md Wahiduzzaman;Rukunuzzaman, Md;Nahid, Khan Lamia;Dey, Bishnu Pada;Sayeed, Maimuna;Rahman, AZM Raihanur;Fathema, Kaniz;Khadga, Mukesh
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • 제25권5호
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    • pp.413-421
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    • 2022
  • Purpose: Perinatal cytomegalovirus (CMV) infection can lead to biliary atresia (BA) in different entities. This study aimed to compare the clinical, hematological, biochemical, and histological features of infants with BA based on their CMV immunoglobulin M (IgM) status at presentation. Methods: This cross-sectional descriptive study was carried out between January 2019 and June 2020 at the Department of Pediatric Gastroenterology and Nutrition at the Bangabandhu Sheikh Mujib Medical University (BSMMU) in Dhaka. Forty-three patients with BA were selected purposively and categorized into either the CMV IgM-positive or CMV IgM-negative BA group. Categorical variables were compared using Fisher's exact test and chi-square tests, while the Student's t-test and Mann-Whitney U-test were used to compare continuous variables. For all statistical tests, a p-value <0.05 was considered statistically significant. Results: Thirty-three (76.7%) of the cases were between 2 and 3 months of age on admission. The clinical, hematological, and biochemical parameters did not differ significantly between the CMV IgM-positive and CMV IgM-negative BA groups. Most (50.0%) of the CMV IgM-positive cases had fibrosis stage F2, while 43.5% of the CMV IgM-negative cases had fibrosis stage F3, with no significant difference between the groups (p=0.391). Conclusion: Our data shows no significant distinction between CMV IgM-positive and CMV IgM-negative BA, suggesting that CMV does not contribute to BA pathogenesis.

능성어(Epinephelus septemfasciatus) immunoglobulin M에 대한 단클론 항체 생산 (Production of monoclonal antibodies to immunoglobulin M of sevenband grouper (Epinephelus septemfasciatus))

  • 김시우;김종오;공경희;오명주;김위식
    • 한국어병학회지
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    • 제34권1호
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    • pp.111-115
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    • 2021
  • Immunoglobulin M (IgM) of sevenband grouper (Epinephelus septemfasciatus) was purified by mannan-binding protein (MBP) affinity column. The purified IgM had an apparent molecular weights of 76 (heavy chain) and 28 (light chain) kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Eight hybridoma clones secreting monoclonal antibodies (mAbs) against sevenband grouper IgM were established. Antibody detection enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the 8 mAbs revealed that optical density (OD) values were clearly different between sera from BSA-immunization and non-immunization of sevenband grouper. These results suggest that the produced mAbs in this study are specifically reacted with IgM of sevenband grouper.

넙치(Paralichthys olivaceus)의 immunoglobulin M에 대한 단클론 항체 생산 (Production of Monoclonal Antibodies Against the Immunoglobulin M of Olive Flounder Paralichthys Olivaceus)

  • 김위식;김기홍;김춘섭;오명주
    • 한국수산과학회지
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    • 제50권2호
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    • pp.169-174
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    • 2017
  • Immunoglobulin M (IgM) was purified from olive flounder Paralichthys olivaceus sera using mannan-binding protein (MBP) and protein L affinity columns (designated as MBPIgM and ProLIgM, respectively). A monoclonal antibody (MAb) against olive flounder IgM was produced. The MBPIgM and ProLIgM had apparent molecular weights of 77, 73, and 28 kDa in SDS-PAGE. Nine hybridomas secreting MAbs against olive flounder IgM were established: five MAbs for MBPIgM (1, 2, 3, 4, and 5) and four for ProLIgM (6, 7, 8, and 9). Western blotting indicated that seven MAbs recognized heavy (H; MAbs 1, 2, 3, 4, 5, 6, and 7) chains and one recognized light (L; MAb 9) chains of IgM, while MAb 8 did not recognize IgM. The results of enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the nine MAbs revealed that the optical density (OD) values of sera differed significantly between BSA- and non-immunized fish, despite some sera from non-immunized fish with slight high OD values. These results suggest that the MAbs produced in this study reacted specifically with the IgM from olive flounder.

SMB(Simulated Moving Bed)를 이용한 IgY(Immunoglobulin Yolk) 분리의 전산모사 (Simulation of IgY(Immunoglobulin Yolk) Purification by SMB(Simulated Moving Bed))

  • 송성문;김인호
    • Korean Chemical Engineering Research
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    • 제49권6호
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    • pp.798-803
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    • 2011
  • 난황에 포함된 IgY는 포유동물에 있는 바이러스나 항원에 반응하는 항체와 같은 역할을 한다. IgY 분리를 위해 난황을 전처리한 후 3-zone SMB를 이용하여 지질단백질들로 부터 분리하는 전산모사연구를 수행하였다. 회분식 크로마토그래피와 pulse input method(PIM) 실험을 이용하여 전산모사 매개변수와 흡착 등온식을 얻었다. Aspen simulator를 이용하여 전산모사를 수행하여 IgY를 분리할 수 있는 SMB 운전조건을 다음과 같이 제시할 수 있었다. 삼각형 이론의 $m_2$$m_3$ 값은 각각 0.18, 1.0이고 스위칭 타임은 419 초이었다. 전산모사 결과 raffinate의 IgY 순도가 98.39%이고 두번째 싸이클에서 순도가 정상상태에 도달하였다.

양이온 계면활성제로 형성된 역미셀을 이용한 초유 Immunoglobulin G의 분리 (Separation of Immunoglobulin G from Colostrum by Reverse Micelles of Cationic Surfactant)

  • 노경현;임지영
    • 한국축산식품학회지
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    • 제24권1호
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    • pp.80-85
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    • 2004
  • 본 연구에서는 역미셀을 이용한 단백질 추출공정에서 적극적으로 활용되지 못했던 양이온 계면활성제에 의한 단백질 추출 가능성을 제시하였으며 초유로부터 IgG의 분리를 위한 반응조건을 조사하였다. IgG의 분리에 적합한 조건은 반응 수용액상의 경우 pH 8, 50 mM KCl었으며 유기용매상의 계면활성제(CDAB) 농도는 100 mM로 나타났다. 위의 조건에서는 초기시료에 존재하는 IgG의 90%이상이 회수되었으며 회수된 단백질의 93%가 IgG로 나타났다. 또한 본 연구는 기존의 역미셀을 이용한 일반적인 단백질 추출공정인 정추출 및 역추출 공정을 이용하지 않고 정추출 공정만을 이용함으로써 추출과정을 단순화하였다.

IgY (Immunoglobulin Yolk) 분리를 위한 3-영역 SMB (Simulated Moving Bed)와 4-영역 SMB 비교전산모사 (Comparative Simulation of 3-zone SMB (Simulated Moving Bed) and 4-zone SMB for IgY (Immunoglobulin Yolk) Purification)

  • 윤상희;김인호
    • Korean Chemical Engineering Research
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    • 제50권5호
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    • pp.866-873
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    • 2012
  • 난황에 포함된 IgY는 포유동물에 있는 바이러스나 항원에 반응하는 항체와 같은 역할을 한다. 난황을 전처리한 후 3-zone와 4-zone SMB를 이용하여 지질단백질들로 부터 IgY를 분리하는 전산모사연구를 수행하였다. 회분식 크로마토그래피에서 전산모사 매개변수와 흡착 등온식 SMB 전산모사 변수를 얻었다. Aspen simulator를 이용하여 전산모사를 수행하여 IgY를 분리할 수 있는 3-zone과 4-zone SMB 운전조건을 비교하여 다음과 같은 결과를 얻었다. IgY와 다른 단백질의 농도와 순도를 모두 고려할 때, 꼭지점인 좌표($m_2$, $m_3$=0.1, 1.1)에서 3-zone SMB가 최적의 조건으로 생각된다. IgY 만을 고려하면 4-zone SMB가 좌표($m_2$, $m_3$=0.06, 0.5)에서 가장 높게 IgY를 분리할 수 있었다. recycle이 없는 3-zone SMB는 꼭지점 좌표에서 좌표이동이 extract의 지질 단백질 농도에 큰 영향을 주었다.