• Title/Summary/Keyword: immunoglobulin M(IgM)

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

  • Song, Sung Moon;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.659-665
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    • 2012
  • IgY (Immunoglobulin Yolk) in egg yolk corresponds to IgG (Immunoglobulin G) in animal serum and plays an important role as immunological proteins in intestines. Carrageenan and Arabic gum were used as pretreatment agents to purify IgY from fresh egg yolk. DEAE (Diethylaminoethyl) Sepharose column in FPLC (Fast Protein Liquid chromatography) was an ion exchange tool to remove contaminants as well as to elute IgY from the column. GF HPLC (Gel Filtration High Performance Liquid Chromatography) enables to measure the molecular weights of IgY and to identify the purified IgY by comparing the molecular weight of standard IgY with the purified one. IgY is a heterogeneous group of different molecular weight and ionic properties, which was investigated with various IE HPLC (Ion Exchange High Performance Liquid Chromatography) columns such as AX, CX and SCX. Three peaks of IgY were separated in the AX column under the conditions of 0.5 M NaCl and pH=8. The SCX column also gave the three peaks of IgY at 0.5 M NaCl and pH=5.

The Comparative Study of IgG, IgM, and IgA in Laboratory Animal Administrated Red-ginseng, Using Immunoglobulin Productivity Assay (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))

  • Lee, Beom-Jun;So, Hyung-Jin;Kim, Jae-Wan;Lew, Jae-Hwan
    • The Journal of Internal Korean Medicine
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    • v.28 no.4
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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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Structural Characterization of an IgM-like Immunoglobulin in the Serum of Swamp Eel, Monopterus albus (드렁허리(Monopterus albus) 혈청내 lgM유사 면역글로불린의 구조적 특성)

  • 손영종;이정우;장정순
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.103-107
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    • 1990
  • lgM-like immunoglobulin was purified from the immune serum of M albus which immunized with bovine Serum albumin(BSA) as an antigen(Ag) and characterized. The Ag-specific antibody activity of the immune serum was increased after the immunization. The purified lgM-like immunoglobulin had a tetrameric structure which had a molecular weight of 800 kD and the monomer of IgM-like Ig had a mass of 199 kD which was composed of two heavy chains (Mol. wt. 70 kD) and iwo light chains (Mol. wt. 29.5 kD). The IgM-like Ig showed hemaggluti nating activity to mammalian RBC slightly.

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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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    • v.56 no.7
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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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    • v.25 no.5
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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.

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

  • Kim, Si-Woo;Kim, Jong-Oh;Kong, Kyoung-Hui;Oh, Myung-Joo;Kim, Wi-Sik
    • Journal of fish pathology
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    • v.34 no.1
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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.

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

  • Kim, Wi-Sik;Kim, Ki-Hong;Kim, Choon-sup;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.2
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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.

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

  • Song, Sung-Moon;Kim, In-Ho
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.798-803
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    • 2011
  • IgY(Immunoglobulin Yolk) is a specific antibody in egg yolk, and it protects human body from virus and antigen. There are a lot of egg yolk components such as lipoprotein and protein. To separate IgY, HPLC(High Performance Liquid Chromatography) and precipitation were used in a batch mode and SMB(Simulated Moving Bed) was adopted for continuous purification of yolk proteins. IgY and other proteins in yolk were separated by using three-zone SMB chromatography. Before performing SMB experiments, batch chromatography and PIM(pulse input method) were performed to find operation parameters and adsorption isotherms. The results of batch chromatography were compared with simulated results using Aspen chromatography. To find the most suitable separation condition in SMB chromatography, simulations in $m_2$-$m_3$ plane on the triangle theory were carried out. $m_2$ = 0.18, $m_3$ = 1.0 and ${\Delta}$t = 419 s are the best conditions for the highest purity of IgY. With this operating parameters(flow rate in three zone and switching time), the purity of raffinate results in 98.39% from Aspen chromatography simulation. Most of the simulation reached steadystate within second recycle.

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

  • 노경현;임지영
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.80-85
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    • 2004
  • This study was carried out to separate immunoglobulin G(IgG) from colostrum using reverse micellar extraction of cationic surfactant and to suggest suitable extraction conditions. The reconstituted colostrum powder was solubilized into a reverse micellar phase containing CDAB(cetyldimethylethyl ammonium bromide) by mixing equal volume of the aqueous and organic phase with constant stirring. The solubilization of proteins from the aqueous to the organic phase was manipulated by pH and ionic strength of the aqueous phase and concentration of surfactant in the organic phase. Based on the SDS-PAGE and densitometry, about more than 90% of initial IgG was remained in the aqueous phase after reverse micellar extraction. Although the aqueous phase contained lactoferrin and bovine serum albumin as minor components, about 93% of the total protein was IgG. The efficient extraction was achieved by the reaction of sodium phosphate buffer(pH 8) containing 50 mM KCl and organic phase containing 100 mM CDAB. The separation of IgG using reverse micellar extraction was simple, highly efficient and easy to be scaled up.

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

  • Yun, Sang-Hee;Kim, In-Ho
    • Korean Chemical Engineering Research
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    • v.50 no.5
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    • pp.866-873
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    • 2012
  • IgY (Immunoglobulin Yolk) is a specific antibody in egg yolk, and it protects human body from virus and antigen. There are a lot of egg yolk components such as lipoprotein and protein. To separate IgY, HPLC (High Performance Liquid Chromatography) and precipitation were used in a batch mode and SMB (Simulated Moving Bed) was adopted for continuous purification of yolk proteins. IgY and other proteins in yolk were separated by using three-zone and four-zone SMB chromatography. Before performing SMB experiments, batch chromatography simulation parameters and adsorption isotherms were obtained. The parameters of batch chromatography were used to simulate SMB using Aspen chromatography. To compare three-zone and four-zone SMB chromatography, simulations in $m_2-m_3$ plane on the triangle theory were carried out. In terms of concentration and purity of both IgY and other lipoproteins, 3-zone SMB process is considered as ideal at the vertex of triangle ($m_2$, $m_3$=0.1, 1.1). 4-zone SMB yields the highest IgY purity at the coordinate ($m_2$, $m_3$=0.06, 0.5), which is the pure raffinate region. In 3-zone SMB without recycle, other lipoproteins in extract are largely affected in purity by small shift from the vertex of triangle ($m_2$, $m_3$=0.1, 1.1).