Name: sampling human indigenous saliva peptidome using a lollipop-like ultrafiltration probe: simplify and enhance peptide detection for clinical mass spectrometry
Uploaded: 2012-08-07T13:00:00
Description: Watch this Scientific Journal Video about Sampling Human Indigenous Saliva Peptidome Using a Lollipop-Like Ultrafiltration Probe: Simplify and Enhance Peptide Detection for Clinical Mass Spectrometry

이 작품은 건강 보조금의 국립 연구소 (R01-AI067395-01, R21-R022754-01, 그리고 R21-I58002-01)에 의해 지원되었다. 우리는 원고의 비판적 읽기를 위해 C. Niemeyer 감사드립니다.

1. LLUF 프로브의 창조 <ol><li> polyethersulfone의 점막 (2cm 2) 충격기의 테두리에 에폭시와 gluing의 점막에 의해 삼각형 폴리 프로필렌의 심폐 (캘리포니아 대학, 샌디에고)의 맹세했습니다. 30 kDa의 분자량 절단 (MWCO)와 부정 청구 polyethersulfone 막가 사용되었습니다. </li><li> LLUF 프로브는 20 ML의 주사기에 연결된 수 있도록 테플론 플루오르 에틸렌 프로필렌 튜브 (내경 / 외경, 0.35/0.50 ㎝) 삼각형 폴?...

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우리는 많은 펩타이드 조각은 인간의 소화되지 않은 타액에 존재하는 것으로 확인되었습니다. 이러한 펩타이드 조각은 프롤린 풍부한 단백질, actin, 알파 아밀라아제, 알파 1 globin, 베타 globin, histain 1, 케라틴 1, mucin 7, 고분자 면역 글로불린 수용체, satherin, S100A9 다양한 형태의 파생 상품입니다. 불확 실한 절단 사이트와 펩티드의 생산에 기여 여러 가지 요인이있을 수 있습니다. 예를 들어, 일부 펩...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> Polyethersulfone의 점막 </td><td> 막 공사 </td><td></td><td> 30 kDa MWCO </td></tr><tr><td> 테플론 플루오르 에틸렌 프로필렌 튜브 </td><td> Upchurch 과학 </td><td></td><td></td></tr><tr><td> 블루 dextran </td><td> 시그마 </td><td></td><td></td></tr><tr><td> 나노 LC 시스템 </td><td> Eksigent </td><td></td><td></td></tr><tr><td> C18 트랩 칼럼 </td><td> 애질런트 </td><td> 5065-9913 </td><td></td></tr><tr><td> LTQ 선형 이온 트랩 질량 분석기 </td><td> 써모 피셔 </td><td></td><td></td></tr><tr><td> 마법사 2 </td><td> 현자-N Research </td><td></td><td></td></tr><tr><td> Acetonitrile-0.1 % 개미의 산성 </td><td> JT 베이커 </td><td> 9832-03 </td><td> LC / MS 학년 </td></tr><tr><td> 물 0.1 % 개미의 산성 </td><td> JT 베이커 </td><td> 9834-03 </td><td> LC / MS 학년 </td></tr></tbody></table>

sampling human indigenous saliva peptidome using a lollipop-like ultrafiltration probe: simplify and enhance peptide detection for clinical mass spectrometry

인간의 타액 프로테옴 및 peptidome가 1-2 공개되어 있지만 그들은 majorly tryptic 타액 단백질의 소화에서 발견되었다. 인간의 타액에있는 원시 펩티드는 질병을 진단하는 질병의 진행을 예측하고, 치료 효능을 모니터링을위한 잠재적인 가치를 제공하기 때문에 외인성 효소와 사전 소화하지 않고 인간의 타액의 토착 peptidome의 확인은 필수적이된다. 적절한 샘플링은 인간 원주민 침이 peptidome의 식별 향상을위한 중요한 단계입니다. 3-4 파편을 제거하는 원심 분리를 포함한 인간의 타액을 샘플링의 전통적인 방법은 너무 시간이 많이 걸리는 임상 사용을위한 적용하는 것입니다. 또한, 원심 분리에 의한 잔해 제거는 감염된 병원균의 대부분을 청소하고 종종 낮은 풍요의 peptidome의 식별을 방해 높은 풍부한 단백질을 제거하지 못할 수 있습니다. 종래의 proteomic 접근법 해당 PRI인 겔 소화와 활용에 marily 활용 2 차원 겔 전기 영동 (2-DE)이 젤 많은 타액 단백질에게 5-6를 식별하는 능력이있다. 그러나,이 접근 방법은 일반적으로 낮은 풍요의 펩티드 / 단백질을 감지할 충분히 구분하지 않습니다. 액체 크로마 토그래피 - 질량 분석계 (LC-MS) 기반 proteomics은 사전 2-DE 분​​리하지 않고 단백질을 식별할 수있는 대안입니다. 이 방법은 높은 감도를 제공하지만, 그것은 일반적으로 사전에 샘플을 미리 분별화 7 어렵게 임상 사용하기 위해 만들어 트립신과 미리 소화 필요합니다. 샘플 준비로 인한 질량 분광법에 지장을 회피하기 위해, 우리는 모세관 ultrafiltration (CUF) 프로브 8-11이라는 기술을 개발했습니다. 저희 연구실에서 데이터 CUF 프로브는 역동적이고 최소한 침략적 태도 8의 동물의 다양한 microenvironments에서 생체내의 단백질을 캡처하는 능력이있다 는걸 증명 -11. 부정적인 압력이 샘플 수집하는 동안 철수 단순히 주사기에 의해 만들어진 이후로 원심 분리가 필요하지 않습니다. LC-MS와 결합 CUF 프로브를 성공적 tryptic - 소화 단백질에게 8-11을 확인했습니다. 본 연구에서는 우리가 쉽게 사람의 구강에 맞을 수있는 롤리팝 같은 ultrafiltration (LLUF) 프로브를 만들어 ultrafiltration 샘플링 기법을 업그레 이드. 트립신의 소화없이 LC-MS에 의한 직접적인 분석은 indigenously 인간 타액은 다양한 단백질에서 파생된 많은 펩타이드 조각을 포함했다. LLUF 프로브와 타액을 샘플링하면 원심 분리를 피할 수 있지만 효과적으로 많은 크고 높은 풍부한 단백질을 제거했습니다. 우리 대량 spectrometric 결과는 많은 낮은 풍요로움의 펩티드가 LLUF 프로브로 큰 단백질을 걸러 후 감지 된 그림. 낮은 풍요의 타액 펩티드가 탐지 크로마 토그래피로 여러 단계의 시료 분리 독립했습니다. 임상 응용의 경우 LLUF 프로브가 통합LC-MS와 D는 잠재적으로 타액에서 질병의 진행을 모니터링하는 미래에 사용될 수 있습니다.

Watch this Scientific Journal Video about Sampling Human Indigenous Saliva Peptidome Using a Lollipop-Like Ultrafiltration Probe: Simplify and Enhance Peptide Detection for Clinical Mass Spectrometry 

Sampling Human Indigenous Saliva Peptidome Using a Lollipop-Like Ultrafiltration Probe: Simplify and Enhance Peptide Detection for Clinical Mass Spectrometry

향후 임상 응용을위한 타액 표본 추출을 고려 롤리팝 같은 ultrafiltration (LLUF) 프로브는 인간의 구강에 맞게 가공되었다. NanoLC-LTQ 질량 분석법에 의한 소화되지 않은 타액의 직접적인 분석은 큰 단백질과 높은 풍부한 단백질을 제거하고 낮은 풍부한 펩티드 더 감지하게 LLUF 프로브의 능력을 보여주었다.

롤리팝 - 같은 Ultrafiltration 프로브를 사용하여 인간의 원주민 타액 Peptidome를 샘플링 : 임상 질량 분광법을위한 펩타이드 감지를 단순화하고 향상

Although human saliva proteome and peptidome have been revealed 1-2 they were majorly identified from tryptic digests of saliva proteins. Identification ...

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