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"Polyclonal antibodies of aflatoxin B1 were successfully produced from New Zealand White female rabbits afterimmunization by the hapten of aflatoxin B1-carboxymethyl hydroxylamine hemihydrochloride (AFB1-CMO)conjugated with bovine serum albumin (BSA) as the antigen. The hapten was synthesized using the carbodiimidemethod with CMO as a linker. Absorption peaks at 362, 264, and 218 nm were observed as a result of characterizationwith UV-Vis spectroscopy, while IR spectroscopy showed peaks at 3448 cm-1 and 1642 cm-1 attributable to thehydroxyl and nitrile groups, respectively. Furthermore, mass spectrometry showed fragmentation at the m/z of 386,368.2, and 310, which confirms that the hapten of AFB1-CMO was successfully synthesized. The hapten was thenconjugated with BSA to serve as an antigen of AFB1 when it was injected into the rabbits. The specificity of the antigentowards its antibody and the confirmation of hapten-BSA conjugation were characterized using the dot blotimmunoassay, which showed a BSA concentration of 1.74 mg/mL. Two weeks after the primary immunization by itsantigen, agar gel precipitation testing showed that the rabbit blood serum had positive results for polyclonal antibodiestagainst AFB1 with the highest concentration of antibodiest of 2.19 mg/mL.Sintesa of Poliklonal Antibodi Aflatoxin B1. Antibodi poliklonal aflatoksin B1 telah berhasil diproduksi pada hewanuji kelinci betina New Zealand White setelah diimunisasi dengan hapten aflatoksin B1-carboxymethyl hydroxylaminehemihydrochloride (AFB1-CMO) yang dikonjugasikan dengan bovin serum albumin (BSA) sebagai antigen. HaptenAFB1 disintesis menggunakan metode karbodiimida dengan CMO sebagai linker. Puncak absorbansi pada 362, 264,218 nm teramati sebagai hasil karakterisasi menggunakan spektrofotometer UV-Visibel, sementara dengan spektrum IRdiperoleh puncak pada 3448.126 cm-1 dan 1642.451 cm-1 yang masing-masing mengindikasikan adanya gugus hidroksildan nitril. Hasil spektrometri massa menunjukkan fragmentasi pada m/z 386, 368.2, dan 310 yang membuktikan haptenAFB1-CMO telah berhasil disintesis. Hapten ini kemudian dikonjugasikan dengan BSA agar dapat berperan sebagaiantigen AFB1 ketika diinjeksikan pada kelinci. Kekhususan antigen aflatoksin B1 terhadap antibodinya dan konfirmasikonjugat hapten-BSA menunjukkan hasil positif pada uji dot blot immunoassay dengan konsentrasi BSA sebesar 1.74mg/mL. Dua pekan setelah imunisasi primer, agar gel precipitation test menunjukkan hasil positif terhadap antibodipoliklonal AFB1 dengan konsentrasi tertinggi sebesar 2.19 mg/mL."

"ABSTRAKAntibodi poliklonal matriks 1 Virus Influenza A H1N1 dapat dimanfaatkan untuk pendeteksian antigen matriks 1 dalam pengembangan sistem diagnostik maupun pengembangan vaksin virus influenza A. Antibodi poliklonal antara lain dapat diperoleh dengan imunisasi kelinci menggunakan antigen rekombinan M1. Protein rekombinan M1 yang digunakan sebagai antigen diekspresikan pada EscherichiacoliBL21 dan dipurifikasi menggunakan resin Ni¬NTA, kemudian digunakan dalam imunisasi 1 ekor kelinci betina, Oryctalogouscuniculus,galur NewZealandWhite. Respon antibodi spesifik M1 diuji dengan ELISA dan westernblot. Hasil uji ELISA yang dinilai pada panjang gelombang 450 nm, menunjukkan titer antibodi yang tinggi pada serum paska imunisasi terhadap antigen M1 rekombinan (0,544) dibandingkan dengan serum pra imunisasi (0,102).Hasil uji westernblotmenunjukkan adanya reaktivitas serum kelinci paska imunisasi dengan pita protein berukuran ~27 kDa, yang diartikan sebagai adanya respon antibodi spesifik terhadap antigen M1, sedangkan serum kelinci pra imunisasi tidak memperlihatkan reaksi dengan pita protein berukuran 27 kDa tersebut. Terlihat pula adanya reaksi non spesifik yang relatif lemah terhadap pita protein lainnya, baik pada serum paska imunisasi maupun pra imunisasi yang menunjukkan adanya residu protein EscherichiacoliBL21 pada sediaan antigen M1 hasil purifikasi. Antibodi poliklonal yang diperoleh dapat digunakan untuk mendeteksi antigen M1 baik untuk pengembangan uji diagnostik maupun vaksin influenza A H1N1.

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ABSTRACTPolyclonal antibody against influenza A H1N1 matrix 1 protein can be utilized for detection of matrix 1 antigen in the development of Influenza A diagnostic system and vaccine. Polyclonal antibody can be obtained by rabbit immunization using M1 recombinant antigen. M1 recombinant proteins that will be used as antigen was expressed in EscherichiacoliBL21 and purified using Ni-NTA resin. This recombinant antigen was used for immunization of female rabbit, Oryctalogouscuniculus,NewZealandWhitestrain. M1-specific antibody responses were tested by ELISA and westernblot. ELISA test results at a wavelength of 450 nm, showed a high antibody titer in the post-immunization serum against the recombinant antigen M1 (0,544) compared with the preimmunization serum (0,102). Westernblottest results showed reactivity of post-immunized serum against a band of ~ 27 kDa protein, which indicate the presence of specific antibody response against M1 antigen, whereas preimmunization rabbit serum showed no reaction with the 27 kDa protein band. The existence of non-specific reactions that are relatively weak against other protein bands was also observed , both in the post-immunization and pre-immunization sera, indicating the presence of residual E.coliprotein in the purified M1 antigen preparation. The Polyclonal antibody obtained in this study can be used to detect M1 antigen, for development of H1N1 Influenza A diagnostic test and vaccine."

"Penelitian bertujuan untuk mendapatkan antibodi poliklonal kelinci yang distimulasi oleh protein rekombinan globular head neuraminidase (NA) dan mengukur titer antibodi poliklonal. Protein rekombinan globular head NA berhasil diekspresikan secara intraseluler pada sel E.coli BL21 codon plus dengan induksi IPTG 0,1 mM dan dipurifikasi menggunakan resin Ni-NTA. Protein rekombinan globular head NA yang telah dipurifikasi digunakan sebagai antigen untuk menstimulasi antibodi poliklonal kelinci. Hasil penelitian menunjukkan bahwa telah dihasilkan antibodi poliklonal terhadap globular head neuraminidase dan titer antibodi paling tinggi dihasilkan sebesar 1,352.

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The aim of this study was to determine rabbit polyclonal antibody stimulated by neuraminidase (NA) globular head recombinant protein and also to measure the polyclonal antibody titer. NA globular head recombinant protein has been expressed in E.coli BL21 codon plus intracellularly induced by 0,1 mM IPTG and has been purified by Ni-NTA resin. The purified of NA globular head recombinant was used as antigen to stimulate rabbit polyclonal antibody.

The result shows that rabbit polyclonal antibody of neuraminidase globular head was produced and the highest antibody titer was 1,352."