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"Providing a comprehensive review of neutralization and the ability to measure and apply antibodies to modern science and medicine, Detection and quantification of antibodies to biopharmaceuticals : practical and applied considerations, provides biotechnology companies and pharma drug development specialists with insight into the design, optimization, and qualification of assays, as well as the establishment of sampling strategies, choice of appropriate assay end-points, and data analysis for the detection and quantification of neutralizing antibodies to biopharmaceuticals. The text also compares the strengths and weaknesses of various assays."

"Applied network security monitoring is the essential guide to becoming an NSM analyst from the ground up. This book takes a fundamental approach to NSM, complete with dozens of real-world examples that teach you the key concepts of NSM.Network security monitoring is based on the principle that prevention eventually fails. In the current threat landscape, no matter how much you try, motivated attackers will eventually find their way into your network. At that point, it is your ability to detect and respond to that intrusion that can be the difference between a small incident and a major disaster.The book follows the three stages of the NSM cycle: collection, detection, and analysis. As you progress through each section, you will have access to insights from seasoned NSM professionals while being introduced to relevant, practical scenarios complete with sample data."

"Translokasi bakteri merupakan kejadian yang diinisiasi oleh adanya reaksi inflamasi pada permukaan usus dan dapat menyebabkan terjadinya sepsis. Bifidobacterium anima/is subspesies lactis merupakan salah satu bakteri probiotik yang dapat memberikan efek anti-inflamasi, sehingga dapat menghambat terjadinya translokasi bakteri. Gen dnaK merupakan sekuens penanda yang dapat digunakan untuk deteksi B. anima/is subsp. lactis. Optimasi dilakukan untuk mendapatkan pasangan primer optimal dalam kuantifikasi B. anima/is subsp. lactis dengan metode kuantitatif Real-time PCR. Isolat DNA diisolasi dari sampel feses bayi menggunakan metode fenol-kloroform. Pasangan primer dirancang berdasarkan sekuen gen dnaK B.ani malis subsp.lacti s [ABOTO1000010.1] menggunakan program pri mer3. Optimasi primer dilakukan menggunakan 5 konsentrasi berbeda, yaitu 50/50, 100/100, 300/300, 500/500, dan 1.000/1.000 nM. Konsentrasi optimal pasangan primer F_HN019_dnaK dan R_HN019_dnaK untuk kurva standar adalah 1.000/ 1.000 nM dengan nilai efisiensi 95.397% dan R2 0,998. Konsentrasi pasangan primer 50/50--1.000/1.000 nM dapat digunakan untuk kuantifikasi DNA target dengan kisaran nilai Ct sebesar 16,13--31,89. Konsentrasi primer dan DNA sampel tidak berpengaruh dan berkorelasi terhadap nilai Ct. Konsentrasi sampel DNA target terkecil yang dapat terkuantifikasi dengan baik oleh pasangan primer F_HN019_dnaK dan R_HN019_dnaK 300/ 300 nM sampai dilusi 10-4 Pasangan primer F_HN019_dnaK dan R_HN019_dnaK dapat dikembangkan untuk kuantifikasi B. anima/is subsp. lactis dalam sampel feses bayi pada kejadian sepsis.

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Bacterial translocation is an event that is initiated by the presence of an inflammatory reaction at the surface of the intestine and can lead to sepsis. Bifzdobacterium anima/is subspecies lactis is a probiotic bacterium that can provide anti-inflammatory effect, so as to prevent the occurrence of bacterial translocation. dnaK is a marker gene sequences that can be used for detection of B. anima/is subsp. lactis. Optimization is performed to obtain optimal primer pair in quantifying B. anima/is subsp. lactis by the method of real-time quantitative PCR. Isolate DNA were isolated from infant feces samples using phenol­ chloroform method. Primer pairs designed based on gene sequences B.anima/is subsp.lactis dnaK f ABOTO1000010.11 using primer3 program. The primary optimization is done using 5 different concentrations, namely 50/50, 100/100, 300/300, 500/500, and 1.000/1.000 nM. F HN019 dnaK optimal primer pair concentrations and R HN019 dnaK for standard curve were 1,000 I 1,000 nM with 95,397% efficiency values and R2 0.998. 50/50--1.000/1.000 nM concentration of primer pairs can be used for quantification of the target DNA with a range of Ct values of 16.13 to 31, 89. Primer concentration and DNA samples have no effect and relation with the Ct value. The smallest concentration of the target DNA sample that can be quantified well by F_HN019_dnaK and R HN019 dnaK primer pair 300/300 nM is up to dilution 10-4 R HN019 dnaK F_HN019_dnaK primer pairs can be developed for the quantification of B. anima/is subsp. lactis in fecal samples of infants on the incidence of sepsis."

"Latar belakang: Dari 36,9 juta orang yang terinfeksi oleh Human Immunodeficiency Virus (HIV) pada akhir tahun 2018 di seluruh dunia, terdapat sekitar 1-2 juta yang terinfeksi HIV-2. Meskipun HIV-2 kurang patogen dibandingkan dengan HIV-1, misdiagnosis infeksi HIV-2 dapat menyebabkan kegagalan pengobatan yang berujung pada perkembangan Acquired Immune Deficiency Syndrome (AIDS). Diagnostik yang akurat diperlukan untuk menentukan apakah suatu individu telah terinfeksi HIV-1, HIV-2 atau koinfeksi HIV-1 dan HIV-2. Metode: Penelitian ini menggunakan DNA sintetik penyandi antigen rekombinan gp125-gp36 HIV-2 yang bersifat immunodominan, lestari, telah dioptimasi kodon untuk sistem ekspresi E. coli, dan telah dianalisis struktur sekunder mRNAnya. DNA sintetik diklona ke plasmid pQE80L untuk diekspresikan, kemudian dipurifikasi menggunakan kromatografi afinitas Ni-NTA. Antigen rekombinan kemudian diuji reaktivitasnya dengan antibodi anti-HIV-2, serta 7 serum positif HIV-1, HBV, HCV, dan serum normal.Hasil: Gen sintetik berhasil dikonstruksi pada plasmid pQE80L dan dapat diekspresikan dengan induksi 0,1 mM IPTG selama 4 jam. Antigen rekombinan terpurifikasi secra optimal pada kondisi denature dengan pH elusi 4,5. Selanjutnya, hasil uji reaktivitas menunjukkan hasil reaktif untuk antibodi anti HIV-2 dan tidak tidak reaktif untuk 7 sampel serum positif HBV, HCV, dan serum normal. Sedangkan untuk serum positif HV-1, terdapat hasil reaktif pada sampel serum nomor 3 yang diduga disebabkan oleh protein kontaminan dari E. coli.Kesimpulan: Antigen rekombinan gp125-gp36 HIV-2 untuk deteksi antibodi anti-HIV-2 telah berhasil dikembangkan, akan tetapi perlu dilakukan optimasi lebih lanjut untuk mendapatkan antigen rekombinan yang benar-benar murni.

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Background: From 36.9 million people worldwide infected by the Human Immunodeficiency Virus (HIV) at the end of 2018, 1-2 million are infected by HIV-2. Although HIV-2 is less patogenic than HIV-1, misdiagnostic of HIV-2 infection could effect to treatment failure which leads to development of Acquired Immune Deficiency Syndrome (AIDS). Accurately diagnostic is required to ascertain whether an individual has been infected HIV-1, HIV-2, or HIV-1 and HIV-2 co-infection.

Method: This study used immunodominant and sustainable synthetic DNA encoding of recombinant antigen gp125-gp36 HIV-2 which was optimized by codon for E. coli expression system, and analyzed the secondary structure of its mRNA. Synthetic DNA was cloned to the pQE80L plasmid to be expressed, purrifyed using Ni-NTA affinity chromatography. Recombinant antigen therefore was verified for reactivity by anti-HIV-2 antibodies and 7 positive serum of HIV-1, HBC, HCV, and normal serum.

Result: The synthetic gene was succesfully constructed on pQE80L plasmid and able to be expressed by induction of 0.1 mM IPTG for 4 hours. Recombinant antigen was optimally purified in denature conditions due to elusion pH of 4.5. Furthermore, the reactivity test revealed reactive result for anti HIV-2 antibodies and unreactive to 7 positive sample serum of HBC, HCV, and normal serum. While positive serum HIV-1 demonstrate a reactive result in sample serum number 3 supposed causing by protein contaminants from E. Coli. Conclusion: Recombinant antigen gp125-gp36 HIV-2 for the detection anti HIV-2 antibodies has been succesfully developed, however further optimization is required in case to obtain truly pure recombinant antigens."

"Expression and reactivity of spike and nucleocapsid recombinant antigens for detection of anti-SARs C0V-2 antibodies

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Expression and reactivity of spike and nucleocapsid recombinant antigens for detection of anti-SARs C0V-2 antibodies"

"ABSTRAKHepatitis C virus HCV menginfeksi lebih dari 170 juta penduduk dunia dan menyebabkan penyakit hati kronis yang berkembang menjadi sirosis dan kanker hati. Diagnosis yang akurat sangat diperlukan untuk memberikan penanganan tepat secara dini, termasuk mencegah penularan virus tersebut secara lebih luas. Pada penelitian ini plasmid pQE80L-HCV_ME telah berhasil dibuat untuk produksi antigen rekombinan HCV. Gen pengkode antigen tersebut dirancang berdasarkan multiepitop yang bersifat imunodominan, lestari, mewakili subtipe HCV di Indonesia dan global. Gen tersebut dibuat dengan teknik DNA sintetik kemudian diklona dari plasmid pUC57 ke pQE80L. Pengklonaan dilakukan menggunakan situs restriksi BamHI dan HindIII dalam sel E. coli Top10. Plasmid pQE80L-HCV_ME kemudian diverifikasi dengan PCR koloni, analisis restriksi, dan sekuensing.

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ABSTRACTHepatitis C virus HCV have been infected more than 170 million people in the world and caused chronic liver disease that lead to liver cirrhosis and hepatocellular carcinoma. Accurate diagnosis is very important to give early proper treatment and to prevent HCV transmission broadly. In this research pQE80L HCV ME plasmid has been successfully created to produce HCV recombinant antigen. Gene that encodes antigen was designed based on multiepitop sequences from immunodominat region, conserve, and represent the most prevalence HCV subtypes in Indonesia and global. The gene was generated through synthetic DNA then be cloned from pUC57 plasmid to pQE80L. Cloning was performed by using BamHI dan HindIII in E. coli Top10 cell. pQE80L HCV ME plasmid then be verified by colonies PCR, restriction analysis, and sequencing."