EFEKTIVITAS IMPLEMENTASI SISTEM MANAJEMEN MUTU ISO 9001:2008 DI MAN 2 MODEL PEKANBARU

Penelitian ini berjudul efektivitas implementasi sistem manajemen mutu ISO 9001:2008 di MAN 2 Model Pekanbaru. Adapun tujuan penelitian ini adalah untuk mengetahui Efektivitas Implementasi Sistem Manajemen Mutu ISO 9001:2008 di MAN 2 Model Pekanbaru. Penelitian ini menggunakan pendekatan kualitatif dengan metode deskriptif. Teknik pengumpulan data yang digunakan adalah observasi, wawancara, dan studi dokumentasi, dengan peneliti sebagai instrument kunci. Teknik pengambilan sampel dalam penelitian ini menggunakan purposive sampling yaitu peneliti menunjuk secara langsung informan yang dianggap dapat memberikan informasi akurat. Uji keabsahan data dilakukan dengan uji reliabilitas dan validitas. Dari hasil analisis deskriptif dengan strategi triangulasi diperoleh pemenuhan klausul ISO 9001:2008 oleh MAN 2 Model Pekanbaru secara garis besar yaitu: 1) Penetapan perencanaan; 2) Pelaksanaan; 3) Pemeriksaan Proses; dan 4) Rencana tindak lanjut. Hasil penelitian ini menunjukkan bahwa implementasi sistem manajemen mutu ISO 9001:2008 sudah berjalan efektif. Hal ini dapat dilihat dari presentase tercapainya target sasaran-sasaran mutu yang telah ditetapkan sebelumnya, dari sembilan target yang ditentukan, hanya ada satu target sasaran mutu yang tidak dapat dicapai, dengan arti sekitar 90% target sasaran mutu dapat tercapai sesuai dengan jangka waktunya. Serta dari rekapitulasi hasil audit internal ditemukan bahwa presentase ketidaksesuaian mengalami penurunan setiap tahunnya. ;---This study, entitled the effectiveness of the implementation of the quality management system ISO 9001: 2008 in MAN 2 Model Pekanbaru. The purpose of this study was to determine the effectiveness of implementation of the Quality Management System ISO 9001: 2008 in MAN 2 Model Pekanbaru. This study used a qualitative approach with descriptive methods. Data collection techniques used were observation, interviews and documentation study, by researchers as a key instrument. The sampling technique in this study using purposive sampling that researchers pointed directly informant who was supposed to provide accurate information. Test the validity of the data is done with reliability and validity testing. Descriptive analysis of the results obtained by triangulation strategy fulfillment of the clauses of ISO 9001: 2008 by MAN 2 Model Pekanbaru outline namely: 1) Establishment of planning; 2) Implementation; 3) Examination Process; and 4) follow-up plan. The results of this study indicate that the implementation of the quality management system ISO 9001: 2008 has been running effectively. It can be seen from the percentage achieving the target goals established quality before, from nine targets set, there is only one target quality objectives can not be achieved, meaning about 90% of target quality objectives can be achieved in accordance with the time period. As well as of the recapitulation of the internal audit found that the percentage of non-conformity has decreased every year

Antibody based affinity capture LC-MS/MS in quantitative determination of proteins in biological matrices

Determination of proteins in complex biological matrices has massive attention and is exploited in many different scientific disciplines. Routinely, proteins are determined using antibody based immuno-metric assays. Although these assays are easy to perform and widely used, interpretation of the results is challenging: cross reactivity, high dose hook-effect, presence of heterophile- or auto-antibodies give rise to false results, sometimes with dramatic consequences. In the quest for more robust assays a combination of antibody sample clean-up, tryptic digestion and mass spectrometric determination is gaining more attention. This review discusses the advantages of antibody based affinity capture and subsequent LC-MS/MS in protein analysis like less false results and possibilities like multiplexing and isoform differentiation. It also considers the interplay between the analytical, biological and biochemical factors, which still give rise to false results, even with mass spectrometry as the ultimate selective detection step. The intention of this review is to point out both strengths and weaknesses of antibody based affinity capture LC-MS/MS in quantitative determination of proteins in biological matrices

Determination of very low-abundance diagnostic proteins in serum using immuno-capture LC-MS-MS

The use of antibodies in “bottom-up “LC-MS workflows to determine low abundant biological active proteins in complex human samples has increased in recent years: immuno-capture analysis combines the workflow of conventional immunological assays with LC–MS analysis. This paper describes typical challenges, such as cross reactivity and the mass spectrometer’s dynamic range, as well as the advantages of isoform differentiation and multiplexing. Additionally, some experimental formats of immuno-capture bottom-up LC–MS analysis of biological active proteins in complex human samples will be discussed

Data-Independent Acquisition for the Orbitrap Q Exactive HF: A Tutorial.

Data-independent acquisition (DIA) is a powerful mass spectrometric technique to perform both protein identification and quantification of complex protein samples. Setting up DIA methods on Orbitrap analyzers requires a thorough overview of the actions the Orbitrap mass spectrometers carry out. This Tutorial is written with the intention to give an overview of the important parameters to consider as well as which measurements to carry out to get the most out of your DIA method when setting it up on an Orbitrap mass analyzer. Instead of giving the optimal DIA settings, all steps in the construction and optimization of the DIA method are shown and discussed in a way that allows tailored DIA methods. They key steps are building the spectral library after sample fractionation, deciding upon the number of data points per chromatographic peak, determining the scan times of each mass spectrometric step, constructing various DIA methods using these data, and evaluating their performance. This proposed DIA method development strategy was tested on digested lysates from Pseudomonas aeruginosa and compared with conventional DDA analysis to put the DIA results into perspective

Instant on-paper protein digestion during blood spot sampling

A concept integrating sampling and protein digestion is introduced here combining fast and simple fabrication by wax printing on filter paper with trypsin immobilized polymer beads. The paper reactors showed promising results with a high degree of protein digestion within fifty minutes in model protein mixtures as well as in human blood. The model protein mixture was used for the evaluation of performance both with and without a reduction and alkylation step. The paper reactors without reduction and alkylation showed between 46% and 75% protein sequence coverage and between five and 20 high confidence peptides (one and five zero missed cleavage peptides, respectively). Compared to a conventional in-solution approach, the paper reactor showed 10% less protein sequence coverage, 29% fewer high confidence peptides and 19% fewer high confidence peptides with zero missed cleavages. Placement of the protein reduction and alkylation step (before or after protein digestion) was shown to be of low importance. The storage stability of the paper reactors with (six weeks) and without (twelve weeks) tryptic peptides was satisfactory. The ability of the paper reactors to digest complex biological samples was investigated by comparison with human whole blood samples prepared using a conventional dried blood spot (DBS) procedure with overnight digestion in non-targeted analysis. The reactors showed a comparable performance with 75 ± 25 for the protein groups compared to 76 ± 5 for the DBS samples. Additionally, 267 ± 72 and 335 ± 11 unique peptides (high confidence) were identified for on-paper digestion and DBS, respectively

Isolation and mass spectrometry analysis of urinary extraexosomal proteins

The aim of the present study was to develop a LC-MS/MS-based proteomic analysis method of urinary exosomal proteins that has the potential to discover disease biomarkers. In short, urinary exosomes from healthy subjects were isolated by immunocapture on magnetic beads, detected by immunofluorescence and TEM, trypsin digested directly on the beads for an accelerated time with no addition of detergents before performing an LC-MS analysis of the trypsinate. To our knowledge, this is the first proteomic analysis of proteins displayed on the outer surface of exosomes. The outer exosome proteome may contain proteins that are of higher biomarker value compared to soluble cargo protein as the proteins projecting into the extracellular milieu might be more directly involved in physiological functions of exosomes. The proteomic analysis identified 49 proteins that were considered significant; the majority is involved in carbohydrate and lipid metabolism or in immune responses. Thirty of the proteins are linked to diseases. The developed proteomic method exploiting urinary exosomes might be of great value in search for diagnostic or prognostic biomarkers of especially metabolic and immune-related diseases

Affinity capture in bottom-up protein analysis – overview of current status of proteolytic peptide capture using antibodies and molecularly imprinted polymers

Antibody-based affinity capture has become the gold standard in sample preparation for determination of low-abundance protein biomarkers in biological matrices prior to liquid chromatography-mass spectrometry (LC-MS) determination. This comprises both capture of intact proteins prior to the digestion step and capture of proteolytic peptides after digestion of the sample. The latter can be performed both using antibodies specifically developed to capture target proteolytic peptides, as well as by the less explored use of anti-protein antibodies to capture the proteolytic epitope peptide. Molecularly imprinted polymers (MIPs), also called plastic antibodies are another affinity-based approach emerging as sample preparation technique in LC-MS based protein biomarker analysis. The current review gives a critical and comprehensive overview of proteolytic peptide capture using antibodies and MIPs in LC-MS based protein biomarker determination during the last five years. The main emphasis is on capture of non-modified peptides, while a brief overview of affinity capture of peptides containing post-translational modifications (PTMs) is provided

Next-Generation Dried Blood Spot Samplers for Protein Analysis: Describing Trypsin-Modified Smart Sampling Paper

This paper describes smart sampling paper to be used for bottom-up protein analysis. Four different manners to immobilize trypsin on cellulose were evaluated. Untreated paper, potassium-periodate-functionalized paper (with and without post-immobilization reduction) and 2-hydroxyethyl methacrylate (HEMA)/2-vinyl-4,4-dimethylazlactone (VDM)-functionalized paper were all used to immobilize trypsin. For the evaluation, Coomassie Brilliant Blue staining of proteins on paper and the BAEE trypsin activity assay needed to be modified. These methods allowed, together with data from mass spectrometric analysis of cytochrome C digestions, us to acquire fundamental insight into protein binding, and trypsin action and activity on paper. All functionalized discs bind more protein than the untreated discs. Protein binding to functionalized discs is based on both adsorption and covalent binding. Trypsin immobilized on potassium-periodate-functionalized discs exhibits the highest trypsin activity when using cytochrome C as substrate. It is proven that it is trypsin attached to paper (and not desorbed trypsin) which is responsible for the enzyme activity. The use of discs on complex biological samples shows that all functionalized discs are able to digest diluted serum; for the best-performing disc, HEMA-VDM functionalized, up to 200 high-confidence proteins are qualified, showing its potential