Conference Proceedings - sipeg.unj.ac.idsipeg.unj.ac.id/repository/upload/artikel/23... · Conference Proceedings !! INTERNATIONAL CONFERENCE ON RESEARCH, IMPLEMENTATION AND EDUCATION

Conference Proceedings

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INTERNATIONAL CONFERENCE ON RESEARCH, IMPLEMENTATION AND EDUCATION

OF MATHEMATICS AND SCIENCES (ICRIEMS) 2014 Yogyakarta, 18 – 20 May 2014

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ISBN 978-979-99314-8-1 !!!!!!!!

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!Global Trends and Issues on Mathematics and Science and The Education

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Faculty of Mathematics and Natural Sciences Yogyakarta State University

!!!ICRIEMS 2014 : Global Trends and Issues on Mathematics and Science and The Education !!!!! Mathematics & Mathematics Education ! Physics & Physics Education!! Chemistry & Chemistry Education ! Biology & Biology Education ! Science Education!!!!!© June 2014 !!!Editorial Board: Hari Sutrisno Wipsar Sunu Brams Dwandaru Kus Prihantoso Krisnawan Denny Darmawan Erfan Priyambodo Evy Yulianti Sabar Nurohman Board of Reviewer Prof. Dr. Saberi bin Othman (Universiti Pendidikan Sultan Idris, Malaysia) Prof. Samsuuk Ekasit (Mahidol University, Thailand) Prof. Dean Zollman (Kansas State University, U.S.) Prof. Tran Vui (Hue University, Vietnam) Prof. Dr. Amy Cutter-Mackenzie (Southern Cross University, Australia) Dr. Alexandra Lynman (Universitas Hindu Indonesia) Prof. Dr. Rusgianto Heri Santoso (Yogyakarta State University) Prof. Dr. Marsigit (Yogyakarta State University) Prof. Dr. Mundilarto (Yogyakarta State University) Prof. Dr. Zuhdan Kun Prasetyo (Yogyakarta State University) Prof. Dr. K.H. Sugijarto (Yogyakarta State University) Prof. Dr. A.K. Prodjosantoso (Yogyakarta State University) Prof. Dr. Djukri (Yogyakarta State University) Prof. Dr. Bambang Subali (Yogyakarta State University)

Proceeding of International Conference On Research, Implementation And Education Of Mathematics And Sciences 2014, Yogyakarta State University, 18-20 May 2014

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Table of Content

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Front Cover Editorial Board and Reviewers Preface Forewords From The Head of Committee Forewords From The Dean of Faculty Table of Content Plenary Session Using Dynamic Visual Representations To Discover Possible Solutions In Solving Real-Life Open-Ended Problems

Prof. Tran Vui Parallel Session MATHEMATICS Probability Density Function of M/G/1 Queues under (0,k) Control Policies: A Special Case

Isnandar Slamet, Ritu Gupta, Narasimaha R. Achuthan !!!!!!-Valued Measure And Some Of Its Properties

Firdaus Ubaidillah , Soeparna Darmawijaya , Ch. Rini Indrati Applied Discriminant Analysis in Market Research

Hery Tri Sutanto Characteristic Of Group Of Matrix 3x3 Modulo P, P A Prime Number

Ibnu Hadi, Yudi Mahatma The Properties Of Group Of 3 3! Matrices Over Integers Modulo Prime Number

Ibnu Hadi, Yudi Mahatma Random Effect Model And Generalized Estimating Equations For Binary Panel Response

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The Implication of Islamic Character Education and Minangkabau Culture to Biological Learning Achivement

Yosi Laila Rahmi

Developing Learning Outcome Based on The Indonesian Qualification Framework Level Six for Biology Education

Zuhdan Kun Prasetyo Analysis of Student’s Misconceptions on Basic Science Concept Through CRI (Certainly of Response Index), Clinical Interview and Concepts Map

Zulfiani Increasing ISTE Program Student’s Activities Using Video on Writing and Retelling

Zulyusri CHEMISTRY The Effects Of Micro- And Nanohydroxyapatite Application In Metal Contaminated Soil On Metal Acccumulation In Ipomoea Aquatica And Soil Metal Bioavailability

Azlan Kamari, Norjan Yusof, Che Fauziah Ishak, Esther Phillip, and Galuh Yuliani

Chromium Extraction from Soil by Using Green Mustard (Brassica juncea)

Tri Santoso, Baharuddin Hamzah, Irwan Said, Ririen Hardani

Biosorption of Technical Direct Dyes by Activated Sludge

Dewi Yuanita Lestari and Endang Widjajanti LFX Effect Activation of Chemical and Physical to Structure and Activated Carbon Quality from Charcoal Obtained Bypyrolysis of Coconut Shell

Djefry Tani, Bambang Setiaji, Wega Trisunaryanti, Akhmad Syoufian

Effects of Calcination Temperatures on Synthesis of LiMn2O4 by Polymer Matrix-Based Alkaline Deposition Method Dyah Purwaningsih, Hari Sutrisno, Dewi Yuanita Lestari

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06 07 08 09 10 11 12 13 14

Silver Nanoparticle Impregnated on The Composite of Bacterial Cellulose-Chitosan-Glycerol as Antibacterial Material Eli Rohaeti, Endang Widjajanti LFX, and Anna Rakhmawati Determination of Glycemic Score of Processed Food from Whole Wheat (Triticum aestivum L.) Flour Dewata’s Variety in Terms of Amylose Content and Starch Digestibility

Febrine Pentadini, Silvia Andini, Sri Hartini, Anik Tri Haryani

Characterization of Quinoline and Quinoline Conjugated Metal as The Base Material of Photodetector

I Gusti Made Sanjaya, Dian Novita and Aldo Swaztyznt Saputra

Characterization of Quinoline and Quinoline Conjugated Metal as The Base Material of Photodetector

I Gusti Made Sanjaya, Dian Novita and Aldo Swaztyznt Saputra

Preparation and Mechanistic Study of ZnO/Zeolite as Catalyst in 1-Pentanol Dehydration

Is Fatimah Effect of Pyrolisis Temperature and Distillation on Character of Coconut Shell Liquid Smoke

Johny Zeth Lombok, Bambang Setiaji, Wega Trisunaryanti, Karna Wijaya

Characterization Chemical Compound Based Pyrolysis Process from Cacao Wastes

Mohammad Wijaya.M Preparation and Characterization of Poly(!-Caprolactone) Microparticle Blends Containing Propranolol HCl and Carbamazepine

Muhaimin, Burkhard Dickenhorst, Roland Bodmeier Production and Characterization of Anti Fim-C Salmonella typhi Native Protein Antibody in Ddy Mice

Muktiningsih Nurjayadi, Umar Hasan, Dea Apriyani, Fera Kurnia Dewi, Irma Ratna Kartika, Fernita Puspasari, Dessy Natalia

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15 16 17 18 19 20 21 22 23 24 25

Synthesis of Star Poly(4-Vinylpyridine) Architecture by Nitroxide Mediated Polymerisation

Nurulsaidah Abdul Rahim, Fabrice Audouin, Johannes G Vos, Andrea Heise

Antifungal Potential Test of Glycoside Compound from Root Woof of Pterospermum subpeltatum C. B. ROB

Pince Salempa, Alfian Noor, Nunuk Hariani, Sudding, Muharram

Test Method Verification of Fe and SiO2 in Industrial Water by Uv-Vis Spectrophotometry at Pt Krakatau Steel Reni Banowati Istiningrum, Intan Permatasari, Idrus Bambang Iryanto Chalcones: The Promising Compounds to Provide New Ways for Cancer Treatment Retno Arianingrum

Electrocoagulation of Detergent Wastewater Using Aluminium Wire Netting Electrode (Awne)

Riyanto and Afifah Hidayatillah Characterization K3PO4/NaZSM-5 Using Xrd and Ftir as a Catalyst to Produce Biodiesel Samik, Ratna Ediati, and Didik Prasetyoko Adsorption Rate Constant and Capacities of Lead(Ii) Removal from Synthetic Wastewater Using Chitosan Silica

Sari Edi Cahyaningrum and Dina kartika Intervention Effect of Liquid Smoke of Pyrolysis Result of Coconut Shell on Profile of pH Fillet of Lates Calcarifer

Sofia Satriani Krisen, Bambang Setiaji, Wega Trisunaryanti, Harno Dwi Pranowo

Phytochemical of Kaempferia Plant and Bioprospecting for Cancer Treatment

Sri Atun Study of Acid Catalysis for Condensationof 4-Hydroxybenzaldehyde With Acetone Sri Handayani Isolation and Identification Secondary Metabolites Compound Ethyl Acetate : N-Hexane (4 : 6) Fraction of Gulma Siam

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26 27 28 29 30 31 32 33 34

Leaves (Chromolaena odorata L.) Sudding

Review of Applications Nanoparticles of TiO2 and ZnO in Sunscreen

Sulistyani A QM/MM Simulation Method Applied to The Solution of Zr4+

in Liquid Ammonia Suwardi, Harno D. Pranowo dan Ria Armunanto

Comparative Study of Methods in The Synthesis of Magnetite (Fe3O4)

Suyanta, Eko Sri Kunarti, Muhamad Muzakir, Citra Pertiwi and Dian Pertiwi

Chemical Constituents of Indonesian Silver Fern (Pityrogramma calomelanos) and Their Citotoxicity

Suyatno, Nurul Hidajati, Khoriyah Umami, and Ika Purnama Sari

Chitosan and N-ALKYL Chitosan as A Heterogeneous Base Catalyst in The Transesterification Reaction of Used Cooking Oil

Tatang Shabur Julianto and Restu Ayu Mumpuni Study on Growth of Carbon Crystal from Charchoal Obtained by Pyrolysis of Coconut Shell

Meytij Jeanne Rampe, Bambang Setiaji, Wega Trisunaryanti, Triyono

Phenolic Compounds from Chloroform Extract of Xylocarpus Moluccensis Stem Bark (Meliaceae)

Tukiran, Nurul Hidayati, Nurul Aini, and Yunita Dwi Setyorahayu

Preparation of Chitin from Shrimp Shells by Papain Latex (Carica Papaya)

Yuli Rohyami, Reni Banowati Istiningrum, Ida Sulistyaningrum

Characterization of Cu(Ii) Complexes of 4-Methylbenzenesulfonylhydrazone and The Potential as Reagent for Phenolic Compound Detection

Yusnita Juahir, Norlaili Abu Bakar, Wan Rusmawati Wan Mahamod, Saripah Salbiah Syed Abdul Azziz, Rozita

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PRODUCTION AND CHARACTERIZATION OF ANTI FIM-C Salmonella typhi NATIVE PROTEIN ANTIBODY IN DDY MICE

Muktiningsih Nurjayadia, Umar Hasana, Dea Apriyania, Fera Kurnia Dewia, Irma Ratna

Kartikaa, Fernita Puspasarib, Dessy Nataliab

aDepartement of Chemistry, Mathematics and Science Faculty, State University of Jakarta, Indonesia

bBiochemistry Division, Department of Chemistry, Mathematics and Science Faculty, Bandung Institute of Technology, Indonesia

Abstract

Typhoid fever is a disease affect many people in developing countries including Indonesia. Prevention can be aided by vaccination to the bacteria; however it still needs further research. This research is aimed to determine the immunogenicity of fim-C native protein Salmonella typhi in vivo as a vaccine candidate in ddY mice. Experimental method is used in this research. This research used antigen of pure fim-C native recombinant protein from its previous research. The ddY mice are categorized into five groups, namely experiment group 1 (protein Fim-C+Freund's Complete/Incomplete Adjuvant), experiment group 2 (protein Fim-C S. typhi), control group 1 (FCA/FIA adjuvants), control group 2 (PBS1x), and normal group (PBS1x). The results showed that the ddY mice produced antibody response after subcutaneous injection of the Fim-C S. typhi native protein +adjuvant or without adjuvant. The antibody responses with Fim-C S. typhi native protein as antigen+adjuvant gave higher absorbans than the Fim-C antigen without adjuvant. It is also showed that the antibody titer from the first to fourth injection has gradually increased. These data showed that Fim-C S. Typhi native protein had higher immunogenicity. It is concluded that the Fim-C S. typhi native protein can be used as a potential recombinant vaccine candidate against typhoid fever. Key words: Salmonella typhi, Typphoi Disease, Fim-C Salmonella typhi native Protein Antibody, Recombinant vaccine

INTRODUCTION Typhoid fever is one of diseases suffered by most people in developing countries, including Indonesia. Typhoid fever is suffered ranging from children to adults. Typhoid fever is easily transmitted to human through food and drink contaminated by Salmonella typhi bacteria in the poor standards of environment hygiene. Typhoid fever happens in Indonesia with an average of 900,000 cases per year. The mortality rate is more than 20,000 cases or 91% of all cases ranging in the age of 3-19 years old. The mortality rate increases in every year [Crump and Mintz, 2010; Verry, 2011]. Thus serious action to prevent and overcome typhoid fever in Indonesia is highly needed. Salmonella typhi bacteria has Fim-C protein, called fimbriae. Fim-C protein, a kind of protein on the surface of bacteria cell, acts as important mediators used for interaction or adharence against host cell [Burrows, 2005; Muktiningsih et al, 2009]. Proteins on the surface of bacteria cell as Fim-C can be used as an antigen; it also induces immune response well [Verma et al.,

Muktiningsih Nurjayadi et.al. / Production And … ISBN. 978-979-99314-8-1

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2009; Toobak et al., 2013; Yang et al., 2013; Moreno et al., 2013]. It is presumed that Fim-C proteins on Salmonella typhi can be potentially used as recombinant vaccine for typhoid fever prevention. Due to its higher level of safety, recombinant vaccine is chosen to be further developed rather than conventional non recombinant vaccine that uses attenuated virus. In the prior research, researchers team from UNJ has succeeded in Fim-C Salmonella typhi gene cloning and sub-cloning on the cloning and expression vectors and its protein purification in both native form and inclusion bodies [Anggraeni R et al, 2012; Pratiwi E et al. 2013; Muktiningsih et al, 2013]. This research is aimed to determine the activity of Fim-C S. typhi recombinant protein in its native form as an antigen in mice ddY through immune response analysis. This information is very important in the development of the potential recombinant Fim-C S. typhi protein as candidate for recombinant vaccine to prevent typhoid fever in human. RESEARCH METHOD A. Production of anti Fim-C S. typhi antibodies Production of anti Fim-C Salmonella typhi native antibodies consisted of preparation stage for mice ddY as animal test, preparation stage for Fim-C S. typhi Native recombinant protein as antigene, immunization and antibodies production, and mice serum/anti-FimC S. typhi antibodies isolation [Harlow and Lane, 1988; Deutsher, 1990; Noer, et al., 1992; Jennings, 1995]. A.1. Preparation stage for mice ddY as animal test Production of anti Fim-C antibodies was conducted in LABTIAP (Agricultural Industry Technology Development and Biomedical Laboratory) BPPT-Serpong. The production of anti-Fim C S. typhi Native antibodies performed on 40 male mice ddY strains, age of 5-6 weeks and weigh of 17-24 grams. Mice were obtained from PT Biofarma Bandung. Mice were maintained in cages placed in the treatment room with temperature condition of 20-240C and 20-70% humidity. Treatment room was made sound-proof and impermeable to keep the air pressure lower than the surrounding so the odor will not come out. Every testing was conducted in different room. 4-8 mice are placed in individual cages of polycarbonate (with stainless steel cover) with size of 41,5x27x15 cm3 each. The individual cages were arranged in 3 level stainless steel racks. During the conditioning process, mice were weighed on day 0, day 3 and day 5. Cage, food, health, and activity checks were also done regularly. The conditioning process is as long as a week. Mice were grouped into three major groups, namely the experiment group (KP), the sick control group (KS) and the normal control group (KN). Experimental group had two sub groups; group immunized by mixture of Fim-C native protein and Frued complete/Incomplete adjuvant (FCA/FIA) which consists of 8 mice (KP-1) and group immunized by Fim-C native protein which consists of 8 mice (KP2). The sick control had two sub groups; group immunized by FCA/FIA which consists of 8 mice (KS-1) and group immunized by Phosphate Buffer Saline (PBS 1x) which consists of 8 mice (KS-2). The normal control group (KN) immunized by Phosphate Buffer Saline (PBS 1x) consists of 4 mice. In the challenge test, only experiment group (KP) and sick control group (KS) were infected by S. typhi bacteria. The normal control group (KN) was not be infected by S. typhi bacteria. After the conditioning process, the researcher took blood from mice’s eyes as much as 1-2 mL as pre immune serum. A day prior to collect pre-immune serum, mice were conditioned to food-fasting.


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A.2. Preparation stage for Fim-C S. typhi Native recombinant protein as antigene As many as 25-100 µg Fim-C S. typhi native protein from previous results was disolved witth PBS 1x bufer in Eppendorf 1,5 mL tube to have a total volume of 100-250 µL. Freund’s complete adjuvant (FCA) or Freund’s incomplete adjuvant (FIA) were added with ratio 1:1. Then performed homogenized mix using vortexs until the mix turned white [Harlow and Lane, 1988; Jenings, 1995]. A.3. Immunization and Antibodies Production Immunization was carried out on the backs of mice in the front section near head subcutaneously as much as 2-3 points for one-time injection. On the first immunization 20 µg Fim C S. typhi native protein that has been prepared as an antigen was mixed with Freund’s complete Adjuvant. One week after the first injection researchers took blood sample from mice’s eyes to prepare the serum. One week after the first injection researchers conducted boosting with 40 g Fim-C mixed with Freund’s incomplete Adjuvant. The second and third boosting were conducted with the 80 g Fim-C after one week of the second and the third injections to obtain the optimal antibody amount [Harlow and Lane, 1988; Noer, 1992]. A.4. Mice serum/anti-FimC S. typhi antibodies isolation Blood sample from mice’s eyes was collected on tubes of centrifuges sterile. Blood sample was incubated at temperature of 370C for 30-60 minutes to a visible separation between serum and platelets. Centrifugation was carried out for 10 minutes with the speed of 5000 g at a 4 oC. Clear liquid/serum was taken out and kept in Eppendorf. Then the serum was stored in –200C [Harlow and Lane, 1988].

B. Characterization of anti Fim-C S. typhi Native antibodies by ELISA Antibodies used in the formation analysis of anti-FimC S. typhi antibodies was mice serum from Bleed I-Bleed IV. Formation analysis of anti-Fim-C protein S. typhi antibodies from day 0 (pre-immune serum) until the 6 weeks was obtained by Enzyme Link Immunosorbant Assay (ELISA).

Antigen (30–300 ng Fim-C S. typhi native protein in50 µL fosfat bufer, 1x PBS) was incubated in microtiter plate wells (IWAKI) in room temperature overnight. Every well was washed 3 times each with 1x PBS containing 1 mM MgCl2 and 0,05% (v/v) Tween-20 (wash buffer). After they were washed, researchers added into 150 µL 5% blotto (5 g skim milk in 100 mL 1x PBS) in every well. And then the microtiter plate was incubated at 370C for 1 hour. After the incubation wells were washed as much as 3 times with wash buffer. As many as 50 µL mice serum (from bleed I/pre-immune serum) - bleed V (fifth weeks), by 100x and 300x dilution were added into each well with prepared ELISA design,and then put them into incubation at 370C for one hour. The microtiter plate wells were washed with wash buffer 3 times. After washing process, 50 µL secondary antibodies was added into each well (anti IgG-mice-HRP dilution 1: 5000 with 1x PBS) [Thermo Scientific Biogen, 2013], and incubated in 370C for 1 hour. After the incubation, the microtiter plate wells were washed with wash buffer 3 times. As much as100 µL substrate TMB (3,3’,5,5’-Tetramethylbenzidine) solution was added into each well and incubated at 370C for 30 minutes (produced blue color). After the incubation, the reaction was stopped by adding 50 µL H2SO4 2M (produced yellow color). Next, the absorbance reading was conducted using ELISA-Reader (Microplate Reader) at wavelength 450 nm [Noer, 1992; Deutsher, 1990; Muktiningsih, 2005; Thermo Scientific, Biogen, 2013]. After the formation of antibodies were known to the maximum researchers collected serum in large


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amount or bleeding terminal as much as 5-10 mL of blood.

RESULT AND DISCUSSION A.Production of anti-Fim-C S. typhi native Antibodies The results of animal conditioning process and pre-immune serum collection process Health, weight, diet, and physical condition checks and observation during the conditioning process showed good result. This was demonstrated by the increase in weight on average 3-5 gram/mice. The observed physical condition of feathers and motion also showed patterns of activity that corresponds to standard conditions. The conditioning place and picture of test animals are presented in figure 1.

Figure 1. The conditioning place and picture of test animals. Mice are grouped into 5 cages.

Every cages containing 8 mice for the experimental groups (KP) and the sick control group (KS) and containing 4 mice for normal group (KN).

The retrieval results of pre-immune serum from sinus orbitalis eyes produced 0,5-1 mL blood/mice and it was 0,2-0,5 mL serum of the total blood samples. Serums were stored in frezer with temperture -20oC for further purposes. The pupose of taking pre-immune serum is as a negative control to the formation of anti Fim-C antibodies. This is aimed to ensure that there is no interaction occurs between Fim-C protein as antigene with mice antibodies before imunization. The production results of anti-FimC S. typhi antibodies Production of anti-FimC S. typhi native antibodies in experimental and control groups was performed in 6 weeks. The process consisted of the first imunization with 20 µg Fim C S. typhi native protein, boosting-1 with 40 µg Fim-C S. typhi native protein. Boosting-2 and boosting-3 were conducted by using 80 µg Fim-C S. typhi native protein. Each step gave a total amount serum 0,2-0,5 mL. After ELISA data was obtained with highest absorbance values, in the sixth week researchers collected blood samples of 4 mice from experimental groups by the maximum as much as 5-10 mL. The blood was prepared to serum containing anti-Fim C S. typhi antibodies as many as 2-4 mL/mice. B. Characterization of anti Fim-C S. typhi native antibodies by ELISA The formation analysis of antibodies used as a primer was taken from mice serum of Bleed I-Bleed IV. ELISA analysis was performed on 4 mice from each group,that are: experimental


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group I (immunized by Fim-C S. typhi native Protein mixed with Freud complete/incomplete adjuvant-KP-1), experimental group 2 (immunized by Fim-C S. typhi native Protein without adjuvant-KP2), control group-1 (immunized by Freud complete/ incomplete adjuvant, KS1), control group-2 (immunized by 1x PBS buffer, KS-2), and normal group (immunized by 1x PBS buffer, KN). The result of development of antibodies formation from 4 mice ddY from each group are presented in table 1. While the analysis results of development formation of anti-Fim C S. typhi antibodies is presented in figure 2 and figure 3. Table 1. The data values of absorbance on development anti Fim-C S. typhi native antibodies

at experimental group (KP) and the sick control group (KS) at antigene concentration 100 ng and 300 ng

bleed to- Experimental

Group- 1 (KP1) Experimental

Group- 2 (KP2) The Sick Control Group-1 (KS1)

The Sick Control Group-2 (KS2)

100 ng 300 ng 100 ng 300 ng 100 ng 300 ng 100 ng 300 ng

bleed -0 0,0285 0,04 0,01125 0,00975 0,048333 0,009333 0,01 0,01

bleed -1 0,0055 0,01825 0,00275 0,008 0,027333 0,009667 0,0165 0,0165

bleed -2 0,19675 0,23275 0,04125 0,09575 0,029 0,037 0,0365 0,0365

bleed -3 0,2715 0,44175 0,19 0,2225 0,042667 0,037667 0,0335 0,0335

bleed -4 0,27925 0,5175 0,1805 0,20275 0,04 0,043 0,047 0,047

Based on the results of analysis on the development of formation anti-Fim C S. typhi antibodies by ELISA shown in Table 1 that Fim-C S. typhi native protein with or whithout adjuvants give satisfied immune response. This is shown by an increase absorbance values from Bleed 0-Bleed 4. The increase of color intensity or absorbance values showed increase in the amount of antibody titer that interacted with Fim-C S.typhi antigene. The blue color was from the oxidation TMB substrate (3,3’,5,5’-Tetramethylbenzidine) to 3,3',5,5'-tetramethylbenzidine diimine by Horse Redish Peroksidase enzyme which bounded to secondary antibodies. Color formation reaction was stopped by adding 50 µL H2SO4 and resulted yellow color and measured at wavelenghts 450 nm [Thermo scientific Biogen, 2013] Data presented in table 1 or figure 2 and figure 3 give information that induction by protein Fim-C S. typhi Native +Adjuvant FCA/FIA as antigene produced higher antibodies than by protein Fim-C S. typhi Native+ PBS 1x. This is in accordance with the literature review stated that the Frued complete/incomplete adjuvant (FCA/FIA) can enhance the formation of immune response [Harlow and Lane, 1988; Fiorino et al, 2012; Moreno et al., 2013]. These results also provide information on specific characters of Fim-C S. typhi native protein that if recombinant protein mixed with adjuvants the resulting immune response will be higher than without adjuvants. The information is also essential that Fim-C S. typhi native protein without adjuvant can also provide a good immune response. This information is very important as the scientific foundation to conclude that the withdrawal of recombinant protein molecule of Fim-C S. typhi native is candidate vaccine recombinant. The results of literature analysis stated that the recombinant vaccine has many advantages. It (1) has higher safety for patients (2) has higher purity (3) generates more spesific immune response (4)


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can be produce in large quantities (5) is easy in production and storage, and some of other advantages [Harlow and Lane, 1988; Fiorino et al, 2012].

Figure 2. Graphic analysis of the formation of anti Fim-C S. typhi native antibodies in

the experimental and the control group. (A) graphic of formation of antibodies in experimental group-1 (KP-1). (B) graphic of formation of antibodies in experimental group-2 (KP2), (C) graphic of control group-1 (KS1), (D) graphic of control group-2 (KS2). The x-axis showed the development of immunization from each bleed 0-4. The y-axis shows the value of absorbance reading of ELISA reader. Blue and red colored diagram shows the amount of protein FimC S. typhi recombinant 100 ng dan 300 ng as antigenes.


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Figure 3. The graphic result of immunogenicity test on antigens Fim-C S. typhi Native

protein. The blue line shows the value of absorbance for a group of mice immunized by Fim-C S. typhi native+Adjuvant FCA/FIA as antigens. The red line shows the value of absorbance for a group of mice immunized by Fim-C S. typhi native+1x PBS as antigens. The green line shows the value of absorbance for a group of mice immunized by Adjuvant FCA/FIA+1x PBS as antigens. The green line shows the value of absorbance for a group of mice immunized by Adjuvant FCA/FIA+1x PBS as antigens. The violet line shows the value of absorbance for a group of mice immunized by 1x PBS as antigens. Condition of ELISA performed on serum dilutiom 100x and secondary antibodies dilution 5000x.

CONCLUSION AND SUGGESTION

Fim-C S, typhi Protein in native form has ben succesfully used as an antigen specific in production of anti Fim-C S. typhi antibodies in vivo in mice ddY. The generated formation of immune response is indicated by the color change of substrate after interaction between Fim-C S. typhi antigen with anti Fim-C S. typhi antibodies. From Bleed 1-Bleed IV, the establishment of spesific anti Fim-C S. typhi antibodies with Fim-C + adjuvant FCA/FIA as antigene gives higher immune response than without adjuvants. The result also provides information that Fim-C S. typhi Typhi protein has good nature of imunogenicity because it is able to make higher response immune without the addition of adjuvant and adjuvant FCA/FIA. So that it can be inferred that Fim-C S. typhi native Protein can be used as vaccine candidates. Significant supporting data about vaccine’s standardized test and clinical trials for human are still needed to make recombinant protein molecules of Fim-C S. typhi native serve as a safe and inexpensive molecular vaccine. The information generated in this study could be made of the scientific basis for the further development of recombinant vaccines for typhoid fever in humans, especially in Indonesia.

-0.16E-16

0.10.2

0.30.40.5

0.60.7

Ab

sorb

ansi

Pengambilan darah Ke-

Grafik Perbandingan Antigen S. typhi Native (300 ng)

Jumlah Antigen Fim-C + Adjuvan

Jumlah Antigen Fim-C + PBS

Jumlah Antigen Adjuvan + PBS

Jumlah Antigen PBS


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Invited article. Emerging Infections. CID 2010:50 (15 january). 241-246 Muktiningsih. 2005. Produk Gen CarA Salmonella Typhi Berukuran 42 kDa yang Dideteksi

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