• Allergy, Asthma & Respiratory Disease
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• v.6 no.6
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• pp.310-314
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• 2018

Purpose: Conventional serum IgE assay was costly, required the skills of expert, and relied heavily on expensive equipment. Quantitative measurement of total IgE using Point of Care Test (POCT) device can be the solution for these limitations. This study evaluated and validated the reproducibility of ImmuneCheck IgE. Methods: This study included 120 patients of allergic diseases such as allergic rhinitis, asthma, drug allergy, food allergy, atopic dermatitis, or anaphylaxis. The reliability of POCT ImmuneCheck IgE was evaluated by comparing results from the naked eye and from the Q-Reader. Intratest reproducibility and intertest correlation were analyzed using intraclass correlation coefficient (ICC). Results: Of the 120 enrolled patients, 51 were males and 69 were females. The ages ranged from 19 to 84 years, with an average age of 51.5 years. The concentration of serum total IgE measured by Phadia ImmunoCAP IgE ranged from 5.95 to 5,000 IU/mL. ICC for Intratest reproducibility of ImmuneCheck IgE by naked eye and by Q-Reader were 0.991 (P< 0.001) and 0.989 (P< 0.001), respectively. In addition, intertest correlation between ImmuneCheck IgE and Phadia ImmunoCAP IgE results of naked eye and Q-Reader were 0.968 (P< 0.001) and 0.948 (P< 0.001), respectively. Conclusion: The ImmuneCheck IgE was reproducible and highly correlated with conventional Phadia ImmunoCAP IgE assay. This result suggests that ImmuneCheck IgE can be a useful tool for rapid and precise detection of total IgE.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.4
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• pp.541-548
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• 2003

The Immunoglobulin $IgG_1$ and $IgG_2$ isotype immune responses of domestic ruminants and mice to Cowdria. ruminantium live infection or by immunization with inactivated organisms were determined by the enzyme linked immunosorbent assay and Western blotting. Immunization of goats with inactivated elementary bodies (IEBs) led to a predominant $IgG_1$ isotype response. This indicated that a Th2 response was induced. After challenge, the IgG isotype responses were mixed whereby both $IgG_1$ and $IgG_2$ antibodies were detected. Two goats that survived virulent challenge had a predominant $IgG_2$ isotype response. In cattle live infection by natur l challenge or experiment led to a predominant $IgG_1$ isotype response. Immunization of cattle with IEBs however led to mixed IgG responses characterized by similar $IgG_1$ and $IgG_2$ ratios. In the mouse live infection led to a predominant $IgG_2$ isotype response. This indicated the mouse developed a true Th1 type cell mediated immune response when inoculated with live organisms. Immunization with inactivated organisms on the other hand led to a dominant $IgG_1$ response. It is evident from this work that the immune responses of ruminants and mice to C. ruminantium are different and that using mice as the experimental model for immune responses to Cowdria ruminantium. is not the appropriate.

• Food Science of Animal Resources
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• v.24 no.2
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• pp.182-188
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• 2004

To determine the concentration of Lactoperoxidase (LPO), an indirect enzyme-linked immunosorbant assay(ELISA) was developed. Anti-LPO egg yolk immunoglobulin(IgY) was transferred to egg yolk by immunizing of Brown hens with LPO. The titer of purified anti-LPO IgY was 1: 520,000. The immunological response of anti- LPO IgY with ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein and lysozyme were evaluated, resulting that the anti-LPO IgY found to be a specific antibody toward LPO and no cross-reaction was observed against ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein, and lysozyme in double immunodiffusion test and ELISA test. In indirect ELISA method, coating concentration of LPO and dilution rate of anti-LPO IgY was 0.25$\mu\textrm{g}$/mL and 1:8,000 respectively. Sensitivity in the standard curve of LPO was ranged from 0.01 to 1$\mu\textrm{g}$/mL using anti-LPO IgY.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7433-7438
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• 2013

Aim: To identify new biomarkers for NPC diagnosis with an anti-EBV Western blot test kit. Methods: Serum samples from 64 NPC patients and healthy subjects with four specific VCA-IgA/EA-IgA profiles were tested with an anti-EBV Western blot test kit from EUROIMMUN AG. Proteins were quantified with scores of intensity visually assigned to the protein bands. The markers which showed statistical differences between the NPC and non-NPC subjects were further evaluated in another 32 NPC patients and 32 controls in comparison with established biomarkers including VCA-IgA, EA-IgA, EBV-related protein IgG, and EBV DNA. Results: Among the markers screened, EA-D p45-IgG showed a statistically significant difference (p < 0.05) between NPC and non-NPC subjects with VCA-IgA positivy. In 32 VCA-IgA positive NPC patients and 32 control subjects, the diagnostic accuracy of EA-D p45-IgG was 78.1% with a positive predictive value of 77.8% and a negative predictive value of 78.6%. In the verification experiment, the specificity and sensitivity of EA-D p45-IgG were 75.0% and 90.6 %, respectively. Conclusions: EA-D p45-IgG might be a potential biomarker for NPC diagnosis, especially among VCA-IgA positive subjects.

• IMMUNE NETWORK
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• v.2 no.4
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• pp.195-201
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• 2002

Background: IgE is closely related to the development of allergies. However, the poor relationship between the specific IgE level and the severity of allergic diseases suggests the possibility of functionally different IgE isoforms. With this in mind, rat basophilic leukemia (RBL)-2H3 activation was analyzed with each type of rat IgE for two parameters, exocytosis and IL-4 mRNA production. RBL-2H3 has been well documented in the rat mucosal mast cell line. Methods: RBL-2H3 cells sensitized with each kind of rat IgE was activated by cross-linking FcRI with B5 (monoclonal anti-rat IgE mouse IgG antibodies). The RBL-2H3 exocytosis was measured by analyzing the ${\beta}$-hexosaminidase level, and the level of IL-4 mRNA synthesis was analyzed using semiquantitative RT-PCR. Rat IgE, which was produced by a parasite infection (REP), was prepared using either Paragonimus westermani metacercariae (REP-PW) or Anisakis simplex third stage larvae (REP-AS). A rat IgE prototype of IR162 was prepared by a peritoneal injection of immunocytoma. Results: The level of exocytosis showed a linear relationship with the rat IgE concentration when REP-PW or REP-AS was applied. However, it exhibited a biphasic response with IR162. In addition, the time course of heating at $56^{\circ}C$ illustrated the similarity between REP-PW and REP-AS, which differed from that of IR162. In contrast, the level of IL-4 mRNA synthesis in the RBL-2H3 cells with IR162 was comparable to that of either REP-PW or REP-AS. Conclusion: These results suggest that functionally different rat IgE isoforms exists in RBL-2H3 exocytosis.

• IMMUNE NETWORK
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• v.3 no.2
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• pp.89-95
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• 2003

Background: Unusually high amounts of N region addition and CDR3 length diversity were found in immunoglobulin (Ig) light chain Vk and Jk joins in patients with rheumatoid arthritis (RA). We sought to determine whether this finding is due to excessive activity of the enzyme responsible for N region addition (terminal deoxynucleotidyl transferase [TdT]) in B lineage cells in bone marrow or from positive antigenic selection of B cells with long CDR3 lengths. Methods: We used FACS to isolate $IgM^+/IgD^+$ B cells (predominantly naive) and $IgM^-/IgD^-$ B cells (predominantly class-switched) B cells from peripheral blood of a patient with RA known to have enrichment for long Vk CDR3s and from that of two normal controls. RT-PCR of VkIII transcripts was performed, followed by sequencing of individual cDNA clones. We analyzed the CDR3 lengths and N region additions in 97 clones. Results: There was enrichment for long CDR3 lengths (11 or 12 amino acids) in both $IgM^+/IgD^+$ and $IgM^-/IgD^-$ B cells in RA compared to B cell subsets in the normal controls. The $IgM^+/IgD^+$ B cell subset in RA was markedly enriched for N region addition and was similar to that seen in the $IgM^-/IgD^-$ subset. Conclusion: These data suggest that enrichment for N region addition and long CDR3 lengths in RA may result from unusually high or prolonged activity of TdT in bone marrow.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.87-94
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• 1997

Clonorchis sinearis is a liver fluke which is the most prevalent helminth of humans in Korea. The better diagnostic measure of clonorchiasis is required for its nationwide control program. The present study observed antigenic bands of C. sinensis and reacting immunoglobulins in serum of infected residents. Adult C. sinensis were recovered from experimentally infected rabbits and soluble crude extract of the worms was used as the antigen after supplementation of E-64, a cysteine proteinase inhibitor. SDS- PAGE of the crude antigen resolved more than 20 protein bands between 200 and 14 kDa. The sera of infected humans collected at an endemic village showed specific IgG and IgE antibodies but little IgM and IgA antibodies. The protein bands of 94, 80, 72, 68, 52, 47, 43, 37, 34, and 28-25 kDa strongly reacted with serum Ig(GMA) or IgG antibody and 64, 62, 52, 47,44, 34,28, and 26 kDa bands reacted with serum IgE antibody. However, the 94, 80, 72, 68, 64, 62, 52, 47, and 40 kDa bands of C. sinensis antigen were found non specific. The protein bands of 43, 34, and 28-25 kDa of C. sinensis are primary target molecules of further analysis.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.953-964
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• 1999

본 연구는 세 종류의 Porphyromonas gingivalis(Pg) antigen의 IgG subclass associated recognition을 평가하기 위해 수행했었다. 총 35명의 조기발병형치주질환자중, Pg381에 대한 IgG2항체의 증가를 보이는 5명이 급속진 행형 치주질환자, IgG4의 증가를 보이는 6명의 환자(국소유년형 치주질환자 2명과 급속진행 형 치주질환자 4명), IgG2+4의 증가를 보이는 2명의 급속진행형 치주질환자 그리고 IgG1+2+4의 증가를 보이는 8명의 환자(국소 유년형 치주질환자 2명과 급속진행형 치주질환자 6명)으로 구성된 21명의 환자를 dot immunoblot analysis를 위해 선택했다. 실험에 사용된 정제된 항원은 Pg381에서 추출한 43-kd fimbrilin protein과 lipoplysaccharide(LPS), Pg A7A1-28(ATCC 53977)에서 추출한 capsularpolysaccharide(CPS)였다. Immunoblotting pattern은 IgG4 antibody가 fimbrial antigen에 강력히 반응함을 보여주었다. Fimbriae에 잘 반응하는 몇몇의 IgG4 antibody역시 antigen에 대해 양성반응을 보였다. 대조적으로 IgG2는 CPS antigen을 일차적으로 인식했다. 전부는 아니지만 대부분의 경우, single이나 group화된 IgG subclass는 모두 LPS antigen을 인식하지 못했다. 같은 group에서 염색강도의 개인적인 차이는 증명되었다. 이런 결과는 조기발 병형 치주질환에서 Pg의 fimbriae와 CPS가 immunodominant antigen이 될 수 있음을 제시한다. 더욱이 IgG subclass antibody가 이런 Pg의 immunoglobulin antigen을 선택적으로 인식함을 알았고, 이는 조기발병형치주질환의 병리에 immunodominant antigen과 함께 IgG의 기능적인 역할을 고려해야 함을 제시한다.

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• Journal of the East Asian Society of Dietary Life
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• v.9 no.2
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• pp.200-206
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• 1999

This study was carried out to get a industrial information about a possibility of IgY antibody production, antimicrobial activity and Properties of IgY antibody in egg yolk. After the initial immunization the anti-Salmonella typhimurium IgY antibody level gradually were decreased from firth week to tenth week. On the other hand, the antibody level in the serum were increased from the first week, reaching its peak in the sixth week. Molecular weights of IgY were estimated approximately 72-75KD in a heavy chain and 30-40KD in a light chain by electrophoresis.