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"While significant progress has been made in the past decade, the current understanding of protein aggregation and its consequences is still immature. Aggregation of Therapeutic Proteins provides an up-to-date resource on protein aggregation and its consequences, and available methods to control or slow down the aggregation process. This book also covers an overview on the causes, consequences, characterization, and control of the aggregation of therapeutic proteins. The knowledge in this book will help pharmaceutical scientists in the development of therapeutic proteins, and also instigate further scientific investigations in this area."

"Latar belakang: Metode konvensional untuk mengkonfirmasi infeksi HIV ialah Western blot. Namun, western blot memiliki keterbatasan yaitu kontaminasi dengan antigen selular manusia dan masalah perbedaan genetik di antara subtipe HIV-1 yang menyebabkan hasil indeterminate dan ketidakakuratan diagnosis infeksi HIV-1 subtipe CRF01_AE yang dominan di Indonesia. Pemeriksaan western blot yang tersedia di Indonesia ialah untuk diagnosis infeksi HIV-1/2 dan tidak bersifat spesifik strain. Metodologi: Pada penelitian ini digunakan protein p24 rekombinan sebagai antigen pada western blot. Dilakukan optimasi ekspresi protein p24 rekombinan HIV-1 CRF01_AE pada Escherichia coli BL21CP dan purifikasi serta western blot untuk mendapatkan informasi awal mengenai reaktivitasnya terhadap serum ODHA di Indonesia. Optimasi ekspresi dilakukan terhadap lama waktu induksi, konsentrasi IPTG, dan suhu induksi. Purifikasi dilakukan dengan metode immobilized metal-affinity chromatography (IMAC) dan sistem purifikasi Ni-NTA [Qiagen] pada kondisi native dengan optimasi pada konsentrasi imidazole dalam wash buffer. Hasil: Konfirmasi protein rekombinan dengan western blot menunjukkan bahwa ekspresi dan purifikasi protein p24 rekombinan telah optimal dan reaktif terhadap serum pasien HIV-1 positif di Indonesia. Kesimpulan: Protein p24 rekombinan dari penelitian ini dapat dikembangkan untuk uji diagnostik western blot berdasarkan subtipe CRF01_AE yang dominan di Indonesia.

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Background: Conventional method for confirmation of HIV infection is western blot. However, western blot has limitation of contamination by human cellular antigen and genetic diversity matter among the HIV-1 subtypes that showed indeterminate result and inaccuracy for the diagnosis of HIV-1 subtype CRF01_AE infection predominantly in Indonesia. The western blot available in Indonesia is for diagnosis of HIV-1/2 which is not strain spesific. This research performed the p24 recombinant protein as the antigen in western blot.

Methods: We conducted the optimization in expression of p24 recombinant protein of HIV-1 subtype CRF01_AE in Escherichia coli BL21CP and purification and the confirmation by the western blot to obtain initial information about the reactivity of this recombinant protein with ODHA (people with HIV/AIDS) in Indonesia. Expression optimization administered in the induction time, IPTG consentration used, dan the induction temperature. The purification of the p24 recombinant protein carried with the immobilized metal-affinity chromatography (IMAC) method in Ni-NTA purification system [Qiagen] in native condition with optimization in the imidazole concentration used in the wash buffer.

Result: The confirmation of recombinant protein by western blot showed the expression and purification of p24 recombinant protein has been optimized well and reactive with the Indonesian HIV-1 positive serum patient.

Conclusion: This result indicated the p24 recombinant protein can be applied for the diagnostic assay development based on predominant HIV-1 subtype CRF01_AE in Indonesia."

"Cell in the distress situation, denaturation of proteins may occur, and may also respond by expressing stress proteins. However, such homeostatis effort does not always succeed and even may lead to disease, including cancer. In distress situation also ensue much protein misfolding. Objective: This research were to explain the role of heat shock protein 40 (Hsp40) and HSP20 in pathogenesis of occured oral squamous cell carcinoma (OSSC) patient which realized human papiloma virus (HPV) infection. Material and method: Tissue biopsy frozen section were taken from BOSC and OSCC patients was cut into three part. Parrafin blocks were made from cutting I, which was subsequently stains with HE to ascertain the type of neoplasm. Cutting II was subjected to DNA isolation. The DNA isolation results were subjected to PCR to amplify L1-HPV gene for fixed the HPV stressor. Protein isolation was treated from cutting III, followed with Blottdot test by using anibody monoclonal anti Hsp40 and Hsp 70 and continued with measurement using densitometer to find the concentration of Hsp40 and Hsp70. The collected data were analyzed with F Test (Manova) and discriminant analysis. Result: This experiment showed the differences in concentration of Hsp40 (p<=0,070) and Hsp70 (p<=0,006) between benign oral squamous cell (BOSC) and OSCC patients which realized HPV infection. Conclusion: This experiment proved that OSCC patients which realized HPV infection indicated an up regulated of Hsp70 concentration, so that there was occurs misfolding of the proteins cell. The misfolding was ensue obstacle of apoptosis and to raise cell poliferation which to storm carcinogenesis. An up regulated of Hsp40 was role as co-chaperone."