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Construction and Expression of Immunotoxin Anti EGFRvIII scFv-HPR Conjugate in Pichia pastoris as A Targeted Drug Candidate for Cancer Therapy Yuliawati, Yuliawati; Soejoedono, Retno Damayanti; Fuad, Asrul Muhamad
ANNALES BOGORIENSES Vol 18, No 1 (2014): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/89

Abstract

Epidermal growth factor receptor variant III (EGFRvIII) is a mutant of EGFR lacking 267 amino acids (exon-2 through 7) within its extracellular domain, resulting in the formation of a new epitope as a tumorspecific target. EGFRvIII is commonly found in GBM, breast, ovarian, prostate, and lung carcinomas. Antibodies or part of antibodies (e.g. single chain variable fragment or scFv) with specific activity against this receptor have been developed. These antibodies are internalized into the cell after receptor binding. Ribonucleases can be cytotoxic due to their inherent ability to degrade RNA, therefore causing cell death via inhibition of protein synthesis. The aim of this research is to construct, clone and express an immunotoxin-based conjugate combining an anti-EGFRvIII scFv with a HPRmut (human pancreatic ribonuclease mutant variant) in Pichia pastoris. The anti-EGFRvIII scFv gene was cloned into yeast expression vector pPICZαA and fused with EGFP gene as a marker. The HPRmut gene was subsequently cloned at the C-terminal of the scFv::EGFP fusion, resulting in the scFv::EGFP::HPR fusion construct. The fusion construct was successfully obtained and nucleotide sequence of plasmid was verified. This construct was used to transform Pichia pastoris SMD 1168H. The gene fusion was successfully transformed and expressed in P. pastoris with a transformation efficiency of 102 cfu/μg DNA. The transformed yeasts were screened on agar media containing up to 1000 μg/ml zeocin. Yeast transformants showed green fluorescent due to the expression of EGFP gene. The recombinant proteins have been expressed and secreted from P. pastoris as showed by immunoblotting and SDS-PAGE analyses, then purified by affinity chromatography method. The selected yeast transformant produced at least 15.85 mg of purified protein per liter of yeast culture.
Construction of a DNA Vaccine Using Glycoprotein Gene and Its Expression Towards Increasing Survival Rate of KHV-Infected Common Carp (Cyprinus carpio) Nuryati, Sri; Alimuddin, Alimuddin; Sukenda, Sukenda; Soejoedono, Retno Damayanti; Santika, Ayi; Pasaribu, Fachriyan Hasmi; Sumantadinata, Komar
Jurnal Natur Indonesia Vol 13, No 1 (2010)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (120.942 KB) | DOI: 10.31258/jnat.13.1.47-52

Abstract

Deoxyribonucleic acid (DNA) vaccine has recently been developed as an alternative vaccine against virus infection.This study was the first step of DNA vaccine development to protect cyprinids including common carp (Cyprinuscarpio) and fancy koi (Cyprinus carpio) from KHV (koi herpesvirus) infection in Indonesia. One of KHV glycoproteingenes, i.e. glycoprotein (GP) was ligated with Japanese medaka (Oryzias latipes) â-actin promoter to generatepAct/GP as a DNA vaccine. Fourty fish in body weight of 10-15 g/fish were individually injected by pAct/GP intomuscle in different dosage of 2.5 μg, 7.5 μg and 12.5 μg/100 μl phosphate buffer saline. Total RNA was extractedfrom the 12.5 μg of pAct/GP-injected fish muscle at 24, 48 and 67 hours post-injection to analyze GP expression byRT-PCR method. Potential of pAct/GP as DNA vaccine was examined by injecting KHV into the 30-days-vaccinatedfish. Both of possitive and negative control fish group were not vaccinated. Possitive control fish group wereinjected with KHV, but negative control fish group were not. KHV-challenged fish were reared for 1 month, and thedeath fish were calculated daily. Result of RT-PCR analysis showed that GP gene expression were detected at 3 dpost-injection. Expression of GP in the vaccinated fish groups helped to improve their survival rate after challengedby KHV. All of fish without DNA vaccination had dead 17 days after KHV injection. The results demonstrated thatpAct/GP had high potency to be used as a DNA vaccine against KHV infection in cyprinids.
Fecal Lipid Content, Serum Lipid Profile, and Intra-Abdominal Fat Accumulation in Normal Rats Supplemented with Rice Bran Oil Hana Fitria Navratilova; Evy Damayanthi; Romadhony Ardiansyah; Intan Nur Fitriyana; Retno Damayanti Soejoedono; Maggy Thenawidjaja
Jurnal Gizi dan Pangan Vol. 15 No. 2 (2020)
Publisher : Food and Nutrition Society of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (295.286 KB) | DOI: 10.25182/jgp.2020.15.2.119-124

Abstract

This study aimed to investigate the mechanism of rice bran oil on altering lipid absorption and blood lipid level in normal rat. Male Sprague Dawley rat age 3 months old, weighted 250–300 g were grouped into three groups: control (aquabidest 1 ml), orlistat (2.16 mg / 200 g body weight), and rice bran oil (1.04 mg γ-oryzanol / 200 g body weight). The intervention was given through oral gavage, daily for 28 days. Indicators observed were growth performance, total cholesterol and serum triglyceride levels, lipid levels in feces, and accumulation of intra-abdominal fat. The results showed that the treatment did not significantly affect body weight gain. Fecal lipid levels of orlistat, rice bran oil and control group respectively were 0.19g; 0.17g and 0.13 g (p<0.05), while the percentage of indigestible lipids for orlistat, control and rice bran oil group were 26%, 17% and 13% respectively (p<0.05). Total cholesterol and serum triglyceride levels in rice bran oil group were significantly lower than controls. Rice bran oil did not significantly affect the percentage of total intra-abdominal fat and the weight of the heart and kidney (p>0.05). The intervention of rice bran oil was shown to reduce blood cholesterol and triglyceride levels in normal mice and did not accumulate intra-abdominal fat. The results suggest that rice bran oil might have an effect on blood lipid regulation but not by preventing lipid absorption.
Preparasi Strip Imunokromatografi Koloid Emas untuk Deteksi Cepat Aeromonas hydrophila Suherman .; Retno Damayanti; Agustin Indrawati
Acta VETERINARIA Indonesiana Vol. 8 No. 3 (2020): November 2020
Publisher : IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/avi.8.3.31-39

Abstract

Aeromonas hydrophila merupakan agen penyebab Motile Aeromonas Septicemia (MAS) yang menyebabkan kerugian ekonomi pada industri akuakultur air tawar. Saat ini, metode diagnostik yang tersedia untuk determinasi A. hydrophila membutuhkan waktu lama dan tidak sesuai diaplikasikan di lapangan. Oleh karena itu, dibutuhkan adanya metode lain yang dapat digunakan sebagai diagnostik yang cepat dan aplikatif di lapangan. Tujuan dari penelitian ini adalah membuat alat diagnostik yang mampu digunakan sebagai pendeteksi agen penyebab MAS. Secara singkat, partikel koloid emas berukuran 31,88 nm dibuat melalui reduksi kimia asam kloroaurat dengan natrium sitrat. Koloid emas sebagai detektor dikonjugasikan dengan antibodi poliklonal anti-A. hydrophila 50 μg/ml pada pH 7. Membran nitroselulosa sebagai membran reaksi, ditetesi dengan antibodi poliklonal anti-A. hydrophila 2 mg/ml di garis T dan antibodi goat anti-rabbit IgG 1 mg/ml di garis C. Hasil dari penelitian ini, strip imunokromatografi yang dikembangkan mampu mendeteksi A. hydrophila dengan deteksi minimum 1,2x105 CFU/ml. Strip bersifat spesifik terhadap A. hydrophila, tidak ada reaksi silang yang ditemukan ketika direaksikan dengan bakteri lain. Analisis sampel menggunakan strip hanya membutuhkan waktu 10 menit. Strip imunokromatografi yang dikembangkan dapat mendeteksi A. hydrophila dengan cepat dan memiliki potensi untuk digunakan di lapangan.
MP-8 Lack of Antibody Formation Against Inactivated Avian Influenza Virus in Ducks and Chickens After Intranasally Immunization Okti Nadia Poetri; Retno Damayanti Soejoedodo; Ni Luh Putu Ika Mayasari; Ekowati Handharyani; Novera Nirmalasanti
Hemera Zoa Proceedings of the 20th FAVA & the 15th KIVNAS PDHI 2018
Publisher : Hemera Zoa

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (397.484 KB)

Abstract

Vaccination is one of control strategies implemented in endemic countries such as Egypt and Indonesia [1,2]. Most commercial AI vaccines available in Indonesia are adjuvanted inactivated AI vaccines applied through intramuscular routes. Vaccine application by subcutaneous or intramuscular injection can cause pain and stress in poultry, the route of vaccine through the nasal drip (intranasal) is a more convenient and painless.                However, respiratory applied inactivated influenza is poorly immunogenic. Therefore prior to developing inactivated intranasal vaccine, it is necessary to study  antibody response to inactivated AI virus which exposed through intranasal route. The aim of our research was to determined antibody response of ducks and chickens against avian influenza virus (AIV) subtype H5N1 after intranasally immunization.
AQ-9 Identification of Sumateran Wild Boar Meat (Sus scrofa vittatus) by Restriction Fragment Length Polymorphism (RFLP) Analysis of Cytochrome b Gene Melani Wahyu Adiningsih; Retno Damayanti Soejoedono; Trioso Purnawarman; Hadri Latif; Rahmat Setya Adji; Okti Nadia Poetri; Dwi Desmiyeni Putri
Hemera Zoa Proceedings of the 20th FAVA & the 15th KIVNAS PDHI 2018
Publisher : Hemera Zoa

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (561.42 KB)

Abstract

Sumateran wild boars have been super abundant in Sumateran forest. In Indonesia, this wildlife condition has led to the exploitation for commercial purpose. The high number of Sumateran wild boars population increases wild boar hunting resulting in an abundant availability of wild boar meat in the food market with extremely cheap price. The macroscopic similarity of wild boar meat and beef has prompted the local people to abuse this situation by selling wild boar meat in traditional market as beef. Based on annual record from Cilegon Class II Quarantine Office in 2014, there were nine smuggling cases or a total of 21.556 kg of wild boar meat smuggling effort that were prevented by Cilegon Quarantine officers. The number of food safety concerns related to smuggling of wild boar or counterfeiting beef with wild boar is a very detrimental condition for consumers, especially consumers in traditional markets.The checking of genuineness or validity of food products is an important effort to protect people from consuming unhealthy food and to indicate whether the food is halal or not. Studies of meat detection should be continuously developed as an effort to protect consumers. Genetic method is the most specific and sensitive method to check food ingredients authenticity by detecting the presence of genetic material or deoxyribonucleic acid (DNA). It results from the specific character of the structure of DNA particles and the possibility of using the information included in them. The most frequent loci used for species identificationin phylogenetics and biodiversity studies are mitochondrial cytochrome b (cyt b).Genetic method is the most specific and sensitive tool for analyzing the authenticity of food ingredients in a molecular level by means of detecting the presence of genetic material or deoxyribonucleic acid (DNA). One of the various methods could be used to detect genetic material is polymerase chain reaction (PCR). Specifically, one of such method frequently used in food industry to observe animal derived product fabrication is PCR restriction fragment length polymorphism (RFLP). PCR-RFLP is based on the comparison of the bands profile generated after certain enzymes digest the DNA target. PCR-RFLP is appropriate for meat testing due its ability in exploiting sequence variation in designated DNA region that allows species differentiation even from closely related species through DNA fragment restrictions selected by suitable restriction enzyme. PCR-RFLP is advantageous since it is simple, cheaper, and easier to be adjusted for routine big-scale studies such as surveillance program.
Correlation of electric conductivity values with the dairy milk quality Nova Dilla Yanthi; Syahruddin Said; Anneke Anggraeni; Retno Damayanti; Muladno .
Jurnal Ilmu Ternak dan Veteriner Vol 23, No 2 (2018): JUNE 2018
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (239.201 KB) | DOI: 10.14334/jitv.v23i2.1694

Abstract

Milk, as the prime source of food for mammals, has an electrolyte to replace the loss of body fluid caused by activity or metabolism process. The total electrolyte concentration can be measured based on conductivity value from the nutritional content. Therefore, the parameter of the quality of milk with conductivity value can be a benchmark for quality and selling value of milk, making it simpler to be implemented in the field. The aim of this research is to analyze the relation between electric conductivity (EC) with the content value of cow milk. The milk was taken from 10-30 cows from a farm in Lembang (district of West Bandung), Pengalengan (district of Bandung), Tasikmalaya, Sumedang, Subang, Sukabumi and Bogor of West Java Province. The milk was put in 50 ml of sterile falcon. The Probe EC count-meter CT-3031 was used to measure EC while the quality of milk was measured by Probe MilkoScanTMFT 120 (Foss).  The milk quality is reflected by protein content, Fat, Total Solid (TS), Solid Non-fat (SNF), Lactose, Density, Acidity and Freeze Point Deviation (FPD). The results of this study show that the EC value in the milk gives a very real positive effect (p=<0.01) to Total Solid (TS), Solid Non-fat (SNF), Lactose, and Freeze Point Deviation (FPD). The value of EC also significantly affect (p=<0.05) the value of density in milk. Therefore, the value of EC can be used to predict the quality value of milk.
Implementasi Mesin Penggoreng Mekanis Untuk Meningkatkan Produktivitas Dan Kualitas Kerupuk Amplang Di Ukm Yuan Kartika Retno Damayanti; Sandra Sandra; Bambang Susilo; Yusuf Hendrawan
Journal of Innovation and Applied Technology Vol 6, No 1 (2020)
Publisher : Lembaga Penelitian dan Pengabdian Kepada Masyarakat Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Based on the survey, SMEs need technology improvements for the frying process so that the process can be done semi-mechanically. Mechanical fryers have a capacity of 3-4 kg/process, larger than manual which have a capacity of 2 kg of dough/process, so meets the SME production target of 30 kg/day. The frying process takes 15 minutes, faster than the manual frying process for 20 minutes. The level of comfort and safety in the frying process is also better, because the equipment is equipped with a frame so that workers are not affected by heat due to the burner stove heating process. Production results are smooth and meet the production targets, and the quality of the amplang cracker products is the same both before and after the semi-mechanical fryer is applied.
APPLICATION MACHINE DRYER MECHANICAL FORCED CONVECTION IN THE PROCESS OF DRYING CASSAVA CHIP Retno Damayanti
Journal of Innovation and Applied Technology Vol 2, No 2 (2016)
Publisher : Lembaga Penelitian dan Pengabdian Kepada Masyarakat Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (408.692 KB) | DOI: 10.21776/ub.jiat.2016.002.02.11

Abstract

Implemented Mocaf flour processing on SMEs still has a weakness in the process of drying cassava chips.The drying process is using natural sun drying. In hot conditions in full sun drying takes 2 to 3 days while in the rainy season between November and April the SMEs unproductive because of the delay time of drying, the chip mocaf grow mushrooms with the consequences of the whole production must be discarded. Thus most of the cluster chose not to produce in the rainy season.Application of forced convectionmechanical dryer is an alternative of drying technology in the fermented cassava chips drying process due have a good quality and a longer storability. Forced convectionmechanical dryer is designed with indirect heating system consisting of a heat exchanger unit, the drying chamber, shelves, blower and frames. Cassava chips drying machine capacity is 40-80 kg/batch is divided into 80 shelves. Fuel is LPG. The results of machine performance test showed an effective drying cassava chips is 3.5 hours by the end of the temperature distribution between 31.9 to 47.6 ° C and a water content of 12.1 to 12.9%, with the desired optimum water content of the cassava chips for further processing into flour mocaf is 12-13%. 
Ancaman terhadap Masuknya Virus Penyakit Mulut dan Kuku melalui Daging Ilegal di Perbatasan Darat Indonesia-Malaysia Risma Juniarti Silitonga; Retno Damayanti Soejoedono; Hadri Latif; Etih Sudarnika
Jurnal Sain Veteriner Vol 34, No 2 (2016): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (818.063 KB) | DOI: 10.22146/jsv.11422

Abstract

Penelitian ini bertujuan untuk menganalisis keberadaan daging ilegal di perbatasan darat Indonesia-Malaysia sebagai ancaman risiko masuknya virus PMK ke wilayah Indonesia. Data primer diambil menggunakan teknik pengumpulan pendapat pakar dengan kuisioner, wawancara mendalam (in-depth interview) dan pengamatan langsung di lapang. Data sekunder diperoleh dari publikasi ilmiah dan tulisan atau data yang tidak dipublikasi (statistik, dokumen dan laporan dari instansi berwenang). Penentuan responden secara purposive sampling. Hasil studi menunjukkan bahwa daging ilegal diperkirakan berasal dari berbagai negara termasuk dari negara/zona yang berstatus endemis PMK seperti Semenanjung Malaysia, Thailand, India dan negara/zona yang dinyatakan tidak diketahui oleh responden. Jenis daging ilegal yang masuk ke Entikong berisiko sebagai sumber infeksi PMK seperti daging beku bertulang tanpa limfoglandula dan jeroan beku tanpa limfoglandula. Berdasarkan jalur dan frekuensi pengangkutan, perkiraan volume pemasukan daging ilegal menunjukkan kemungkinan daging masih bisa lolos melalui jalur non-kendaraan. Kondisi-kondisi tersebut mengindikasikan bahwa pemasukan daging ilegal dapat sebagai ancaman risiko masuknya virus PMK ke Indonesia khususnya di perbatasan darat Indonesia-Malaysia, Entikong. Perlu dilakukan upaya pencegahan untuk mengurangi ancaman risiko yaitu dengan melakukan pengawasan yang lebih ketat di pintu-pintu pemasukan dengan koordinasi lintas instansi di perbatasan untuk bersama-sama mencegah pemasukan daging ilegal.
Co-Authors . Darmawi Achmad Agus Priyono Adji, Rahmat Setya Agus Setiyono Agus Wahyudi Agustin Indrawati Anneke Anggraeni Anugrah Nur Rahmanto Asrul Muhamad Fuad Asrul Muhamad Fuad, Asrul Muhamad Assaat, Lusiani Dewi Ayi Santika Bagas Setya Dian Nugraha Bagus Nanang Luwito Bambang Susilo BIBIANA W LAY Dewi Elfidasari Dwi Desmiyeni Putri Ekowati Handharyani Emilia A Erick Teguh Leksono Etih Sudarnika Evy Damayanthi Fachriyan Hasmi Pasaribu Fachriyan Hasmi Pasaribu Fachriyan Hasymi Pasaribu Fajriah, Sofa Fitrianti Rahayu Hadri Latif Hana Fitria Navratilova Handika Martu Kurniawan Hapsari Mahatmi Harry Tetra Antono I Nyoman Suartha I wayan Teguh Wibawan Intan Nur Fitriyana Irvan Faizal Iskandar, Layla Nurina Kartika Ivandini Tribidasari Anggraningrum Ketut Karuni Nyanakumari Natih Komar Sumantadinata Lilis Suryani M. Lutfi Maggy Thenawidjaja Suhartono Makmur iknu wijaya Mangaraja Pidoli Tampubolon Melani Wahyu Adiningsih Mirnawati Sudarwanto Monika Danaparamitha Andriani Muhammad Hambal Natih, Ketut Karuni Nyanakumari Ni Luh Putu Ika Mayasari Nova Dilla Yanthi Novera Nirmalasanti Nurhidayah Nurhidayah Okti Nadia Poetri R.A. Heryani Wahyuningrum Rahmat Setya Adji Ramlah Ramlah Razali Daud Retno Wijayanti Risa Tiuria Risma Juniarti Silitonga Rizky Alviodinasyari Romadhony Ardiansyah Rudy Ariyanto Saepudin, Endang Sandra Sandra Siti Gusti Ningrum, Siti Gusti sri murtini . Sri Nuryati Suherman . Sukenda . Surachmi Setiyaningsih Sutanti, , Syafril Daulay Syahruddin Said TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Trioso Purnawarman Ummu Balqis Usman Nurhasan Wijaya, Makmur Yuliawati Yuliawati Yus Rusila Noor Yus Rusila Noor Yusuf Hendrawan