• KSBB Journal
• /
• 제8권4호
• /
• pp.405-408
• /
• 1993

Absidia zychae NRIC 1199 produced two forms of carboxypeptidase(CPZ-1 and CPZ-2) which were distinguished in their isoelectric points but had almost identical properties(1). The amino acid sequences for the N-terminal of both enzymes were the same (Tyr-Thr-Ser-Pro-Lys-Leu-Xaa-Asp-Pro-Asp-Val) and any significant difference was not observed between amino acid compositions of the two enzymes. The ouchterlony double diffusion technique using antibody raised against the CPZ-2 protein demonstrated a good cross-reaction between CPZ-1 and CPZ-2 Genomic Southern analysis showed only one gene encoding CPZ in the genome of Absidia zychae. However, a significant difference between two enzymes was observed on peptide map using Staphylococcus aureus V8 protease, distinguishable only one band, indicating that multiple forms of CPZ are caused by post-translational modification, such as deamidation.

• 한국동물학회지
• /
• 제35권1호
• /
• pp.17-22
• /
• 1992

The three yolk polypeptides have been isolated and partially characterized. Their molecular weights of YPI, YP2, and YP3 were 48, 000, 47, 000, and 46, 000, respectivelv, as judged by SDS-polyacrvlamide gel electrophoresis. They have different digestion products upon in situ peptide mapping by limited proteolysis. Two-dimensional gel electrophoresis showed that their isoelectric points were heterogeneous from 5.92 to 6.54. And thew showed three different antigen-antibody reactions when each polvpeptides is reacted with antisera made to a mixture of all of three. These data reported here indicate that the yolk proteins are consisted of distinctive polypeptides in Drosophlla sp. (robusta species group).

• Molecules and Cells
• /
• 제44권5호
• /
• pp.328-334
• /
• 2021

The advent of the major histocompatibility complex (MHC) multimer technology has led to a breakthrough in the quantification and analysis of antigen-specific T cells. In particular, this technology has dramatically advanced the measurement and analysis of CD8 T cells and is being applied more widely. In addition, the scope of application of MHC multimer technology is gradually expanding to other T cells such as CD4 T cells, natural killer T cells, and mucosal-associated invariant T cells. MHC multimer technology acts by complementing the T-cell receptor-MHC/peptide complex affinity, which is relatively low compared to antigen-antibody affinity, through a multivalent interaction. The application of MHC multimer technology has expanded to include various functions such as quantification and analysis of antigen-specific T cells, cell sorting, depletion, stimulation to replace antigen-presenting cells, and single-cell classification through DNA barcodes. This review aims to provide the latest knowledge of MHC multimer technology, which is constantly evolving, broaden understanding of this technology, and promote its widespread use.

• 공업화학
• /
• 제35권4호
• /
• pp.335-340
• /
• 2024

본 논문에서는 UIO-66에 항체 기능성을 도입한 유무기 하이브리드 소재를 합성하고 이를 표면 플라즈몬 공명(surface plasmon resonance, SPR) 분석법에 접목하여 옥시토신과 같은 작은 분자를 검출하는 감도를 향상시키고자 하였다. 옥시토신은 암, 알츠하이머, 심부전증 진단에 중요한 생물학적 표지 펩타이드 분자로 알려져 있으며, 이를 수 펨토몰(femtomole, fM) 농도 수준까지 검출하기 위해 다공성이며 표면적이 우수한 metal organic frameworks 중 하나인 UiO-66-(COOH)₂ 소재를 신호증폭용으로 활용하면서 옥시토신에 특이적인 항체 페어를 이용하는 표면 샌드위치 분석법을 개발함으로써 선택성을 향상시키고자 하였다. 이를 위해 먼저 선정한 각 옥시토신 특이적 항체가 옥시토신에 대해 강하게 결합하는지 그리고 각 항체가 옥시토신의 서로 다른 결합사이트에 결합하는지를 실시간 SPR 분석법으로 확인하였다. 선정한 항체 중 한 개(예: anti-OXT [OTI5G4])를 SPR용 금 박막 칩 표면에 고정하고, 옥시토신을 흘려준 후, UiO-66-(COOH)₂에 컨쥬게이션된 다른 항체(예: anti-OXT[4G11])를 순차적으로 흘려주어 표면에 샌드위치 복합체(anti-OXT[OTI5G4]/옥시토신/UiO-66-(COOH)₂-anti-OXT[4G11])를 형성하였을 때 옥시토신 농도에 따라 SPR 신호가 변화하는 것을 실시간으로 모니터링하였다. 그 결과, UiO-66-(COOH)₂를 사용하지 않았을 때 대비 약 백만 배 이상 감도를 증폭시켜 약 10 fM까지 검출 가능함을 보여주었다.

• Journal of Pharmaceutical Investigation
• /
• 제29권2호
• /
• pp.87-92
• /
• 1999

Drug delivery to the brain may be achieved by producing chimeric peptide, attaching the drug to protein 'vectors' which are transported into the brain from the blood by a receptor-mediated transcytosis through the blood-brain barrier (BBB). Since the BBB expresses high concentrations of transferrin receptor, and it was reported that anti-transferrin receptor mouse monoclonal antibody (OX26) undergoes transcytosis through the BBB, it is logical to assume that a drug delivery system via transferrin receptor-mediated transcytosis is a promising strategy. In the present study, therefore, we tested feasibility of several OX26 based vectors for the brain delivery of a model drug. Avidin-based delivery vectors such as OX26-streptavidin (OX26-SA), OX26-neutralite avidin (OX26-NLA) were chemically synthesized vectors and OX26 immunoglobulin G 3 type $C_{H}3$ fusion avidin $(OX26\;IgG3C_H3-AV)$ was genetically engineered. To improve the efficiency of producing chimeric peptide, we used avidin-biotin technology. Pharmacokinetics of $[^3H]biotin$ bound to OX26-SA, OX26-NLA and $OX26\;IgG3C_H3-AV$ was determined by intravenous injection technique, and their stabilities in plasma were analyzed using HPLC. The brain delivery of $[^3H]biotin$ bound to OX26-SA, OX26-NLA and OX26\;$IgG3C_{H}3-AV$ (expressed as %ID/g brain) was $0.22{\pm}0.01$, $0.18{\pm}0.01$ and $0.25{\pm}0.09$, respectively. The areas under the plasma concentration versus time curve (AUC) for OX26-SA, OX26-NLA, $OX26\;IgG3C_H3-AV$ from time zero to 60 min were $209{\pm}10$, $195{\pm}9$, $134{\pm}29\;%ID\;min/ml$ respectively and their total clearances $(CL_{tot})$ were $1.00{\pm}0.09$, $1.08{\pm}0.07$ and $1.54{\pm}0.29\;ml/min/kg$, espectively. These results showed that these vectors possess preferable pharmaceutical (e.g., resonable stability) and pharmacokinetics (e.g., significant brain uptake and enhanced AUC) for brain delivery. Therefore, these vectors may be broadly useful in the brain delivery of drugs that are not transported into the brain to a significant extent.

• Molecules and Cells
• /
• 제40권12호
• /
• pp.954-965
• /
• 2017

Mammalian genomes are well established, and highly conserved regions within odorant receptors that are unique from other G-protein coupled receptors have been identified. Numerous functional studies have focused on specific conserved amino acids motifs; however, not all conserved motifs have been sufficiently characterized. Here, we identified a highly conserved 18 amino acid sequence motif within transmembrane domain seven (CAS-TM7) which was identified by aligning odorant receptor sequences. Next, we investigated the expression pattern and distribution of this conserved amino acid motif among a broad range of odorant receptors. To examine the localization of odorant receptor proteins, we used a sequence-specific peptide antibody against CAS-TM7 which is specific to odorant receptors across species. The specificity of this peptide antibody in recognizing odorant receptors has been confirmed in a heterologous in vitro system and a rat-based in vivo system. The CAS-TM7 odorant receptors localized with distinct patterns at each region of the olfactory epithelium; septum, endoturbinate and ectoturbinate. To our great interests, we found that the CAS-TM7 odorant receptors are primarily localized to the dorsal region of the olfactory bulb, coinciding with olfactory epithelium-based patterns. Also, these odorant receptors were ectopically expressed in the various non-olfactory tissues in an evolutionary constrained manner between human and rats. This study has characterized the expression patterns of odorant receptors containing particular amino acid motif in transmembrane domain 7, and which led to an intriguing possibility that the conserved motif of odorant receptors can play critical roles in other physiological functions as well as olfaction.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
• /
• pp.287-287
• /
• 1994

In the present study, it was aimed to asses the possibility that calcitonin gene-related peptide (CGRP) released in response to transient hypotension may contribute to the reflex autoregulation of cerebral blood flow as a putative modulator. Changes in pial arterial diameter (mean, 33.0 ${\pm}$ 1.1 $\mu\textrm{m}$) with changes in systemic arterial blood pressure (mean, 101.9 ${\pm}$ 2.7 mmHg) were observed directly through a closed cranial window in anesthetized normotensive rats. Image of the pial vessels was captured with a stereoscope connected to a CCD video camera and the diameter was measured with a microscaler. In the capsaicin-treated rats (one day prior to experiment, 50 nmol capsaicin injected intracisternally), both vasodilater and vasoconstrictor responses evoked by a transient hypotension and the reverse of blood pressure were markedly attenuated or almost abolished. When changes in pial arterial diameter were plotted as a function of changes in blood pressure, the slopes of both regression lines (for vasodilators and vasoconstrictors ) were markedly reduced. Similar reductions were evidenced under treatment wi th the CGRP antibody serum (1:1,000) and following CGRP receptor desensitization. However, the autoregulatory mechanics were neither affected by treatment wi th spantide (1 ${\mu}$M), substance P antagonist, nor by substance P receptor desensitization. Suffusion wi th mock cerebrospinal fluid containing CGRP and cromakalim caused a vasodilatation in a concentration-dependent manner, respectively and their effects were antagonized by glibenclamide. Substance P produced a vasodilatation, which was, however, little affected by glibenclamide. These observations indicate that the CGRP released from the perivascular sensory fibers in response to a hypotension is implicated in the modulation of the autoregulation of cerebral blood flow.

• 대한임상검사과학회지
• /
• 제39권1호
• /
• pp.42-48
• /
• 2007

This study was carried out to review and evaluate anti-CCP ELISA assay for diagnostic in RA patients from an early arthritis clinic (EAC). The subjects were obtained from patients visiting the outpatient clinic of the Dept. of Rhumatology med.of P hospital in Daegu, during 6 months from July 1, 2006 to December 31, 2006. The subjects were 140 cases : 80 cases from RA patients (60 women and 20 men; mean age 58 years; range, 32-68 years) confirmed by clinical diagnostic. 39 cases of these RA patients were classified as having early RA (EAC). 50 cases (non-RA) did not fulfill the criteria for RA, and 10 cases were from healthy individuals. We performed the analysis with solid phase-ELISA method (ETI-max3000, Diasorin; Italy) for anti-CCP and Nephelometry assay (Roche/Hitachi 902 analyzer; USA) for RF. The results obtained were summarized as follows ; anti-CCP ELISA is more specific than RF Nephelometry assay (specificity 94% vs 90%) to diagnose RA patients with suspected EAC (early arthritis clinic). The combination test "anti-CCP and RF" had a very high specificity (specificity 98.3%, PPV; RA group 96%, EAC 95%), the difference was statistically significant (p<0.05). Anti-CCP ELISA had more sensitivity in EAC (Early arthritis clinic) patients than chronic RA patients (sensitivity 64% vs 24%, respectively), anti-CCP of RA group and EAC group was more specific than RF (anti-CCP PPV; 92%, 89% vs 89%, 81% respectively), the difference was statistically significant (p<0.05). The difference of antibody concetration between anti-CCP and RF for RA and the control group is statistically significant (p<0.05). In conclusion, anti-CCP ELISA testing may be useful if performed concomitantly with RF Nephelometry assay to diagnose RA patients with suspected EAC (early arthritis clinic).

• Journal of Yeungnam Medical Science
• /
• 제32권2호
• /
• pp.127-131
• /
• 2015

Churg-Strauss syndrome (CSS) is a necrotizing vasculitis with extra-, peri-vascular eosinophilic infiltration. Chronic symmetric polyarthritis with the presence of rheumatoid factor (RF) and anti-cyclic citrullinated peptide antibody are the mainstay of rheumatoid arthritis (RA) diagnosis. Mononeuritis multiplex is a peripheral neuropathy involving more than 2 separate nerve areas. A 62-year-old male patient was referred for left foot drop and polyarthritis of both hands and feet for 4 months. During evaluation, mononeuritis multiplex was detected on nerve conduction study and electromyography tests: vasculitis with neutrophil, eosinophil, and lymphocyte infiltration on peroneal nerve biopsy. A positive response to methacholin and bronchodilator was observed on the pulmonary function test. Radiologic tests showed peri-articular soft tissue swelling and osteopenia on both hand and foot. Marked peripheral eosinophilia, high RF, and positive perinuclear anti-neutrophil cytoplasmic antibody were detected on blood tests. Here, we report on a patient with overlap syndrome of CSS and RA with review of the relevant literature, from which a few references to overlap syndrome of CSS and RA were available.

• Asian-Australasian Journal of Animal Sciences
• /
• 제20권3호
• /
• pp.405-411
• /
• 2007

Gal-13 is an antimicrobial peptide isolated from chicken intestine. Ninety chickens were randomly divided into two groups (45 chickens for each group) to determine the effect of oral administration of Gal-13 on the acquired immune response. The chickens in the first group were fed a diet without Gal-13 as the control, and the chickens in the second group were fed the same diet, except that Gal-13 ($1{\mu}g/ml$) was suspended in drinking water just after hatching. Samples of blood, thymus, bursa of fabricius and spleen were taken at day 1, 4, 7, 10 and 17. The chickens in both groups received infectious bursal disease virus vaccine at day 20, and then sera samples were collected for analysis at 14, 21, 28 and 35 days after vaccination. The results showed: (1) Gal-13 could enhance the content of immunoglobulin (Ig)G at the age of 4 to10 days (p<0.05) and IgM at the age of 4 and 10 days (p<0.05) in the serum; (2) In vitro experiments showed that Gal-13 (0.625-1.250${\mu}g/ml$) enhanced the proliferation of peripheral blood lymphocytes of the chickens stimulated by lipopolysaccharide (LPS) and concanavlin A (ConA). Compared to the control, Gal-13 (1 ${\mu}g/ml$) enhanced the proliferation of bursa lymphocytes at 17 days of age (p<0.01) and thymus lymphocytes at 7 days of age (p<0.01), but restrained lymphocyte proliferation in chicken spleen and differed significantly at day 10 (p<0.01); (3) Gal-13 enhanced infectious bursal disease virus antibody in sera of chickens 21 days after infectious bursal disease virus vaccine administration (p<0.05). These results suggested that Gal-13 could modulate adaptive immune responses of chickens.