Pengaruh Rasio Doxorubisin:Apoferritin terhadap Kapasitas dan Efisiensi Enkapsulasi Doksorubisin dalam Apoferritin

https://doi.org/10.22146/jrekpros.24868

Novita Wiwoho(1*), Maria Christina Prihatiningsih(2), Mujamilah Mujamilah(3), Grace Tj. Sulungbudi(4)

(1) Sekolah Tinggi Teknologi Nuklir
(2) Sekolah Tinggi Teknologi Nuklir-BATAN, Babarsari, Yogyakarta
(3) Pusat Sains dan Teknologi Bahan Maju-BATAN, Kawasan Puspiptek Serpong, Tangerang Selatan
(4) Pusat Sains dan Teknologi Bahan Maju-BATAN, Kawasan Puspiptek Serpong, Tangerang Selatan
(*) Corresponding Author

Abstract


Doxorubicin is a chemotherapy drug which is very toxic and causes many side effects. To reduce side effects, doxorubicin can be encapsulated by apoferritin into apoferritin doxorubicin (Apo-Dox) system. In this research, various mass of doxorubicin i.e. 0.17 mg (S1), 0.26 mg (S2), 0.35 mg (S3), and 0.52 mg (S4) were encapsulated with 21.50 mg apoferritin. Encapsulation process was carried out by lowering pH medium for apoferritin dis-assembly, doxorubicin addition and dialysis for gradual and controlled pH-increase of medium to support re-assembly of apoferritin and doxorubicin encapsulation. End-result samples were then centrifuged and washed to separate the unreacted doxorubicin and apoferritin’s subunits. Doxorubicin encapsulation efficiency was determined using microplate reader spectrophotometry. The highest encapsulation capasity was 3.87 g dox/mg apo for S4 samples. Increasing the weight of doxorubicin gives more significant effect on increasing the reactive weight of apoferritin, which reached 93.73% (S4 sample). Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) analysis confirm that apoferritin is in the Apo-Dox sample.

 

ABSTRAK

Doksorubisin merupakan obat kemoterapi yang sangat toksik dan menimbulkan berbagai efek samping. Untuk mengurangi efek samping tersebut, doksorubisin dapat dibungkus dengan protein apoferritin menjadi Apoferritin-Doksorubisin (Apo-Dox). Dalam penelitian ini, doksorubisin dengan berat 0,174 mg (S1), 0,261 mg (S2), 0,349 mg (S3), dan 0,523 mg (S4) dienkapsulasi apoferritin dengan berat 21,487 mg. Proses enkapsulasi dilakukan dengan mengubah pH larutan melalui penambahan glisin asetat, tris base, dan dialisa dalam tris asetat. Larutan di centrifuge dan di cuci untuk menghilangkan doksorubisin dan sub-unit apoferritin yang tidak bereaksi. Konsentrasi doksorubisin yang terenkapsulasi dalam apoferritin (μg dox/mg apo) ditentukan menggunakan microplate reader spectrophotometry. Konsentrasi cenderung mencapai optimum mulai sampel S2 dan hanya sedikit meningkat untuk sampel S3 dan S4 dengan nilai maksimum pada S4 sebesar 3,87 μg dox/mg apo. Peningkatan jumlah doksorubisin akan meningkatkan jumlah apoferritin yang bereaksi, dan diperoleh nilai efisiensi reaksi apoferritin tertinggi 93,73% untuk sampel S4. SDS-PAGE membuktikan bahwa apoferritin berada pada sampel Apo-Dox.


Keywords


Enkapsulasi; doksorubisin; protein apoferritin; Apo-Dox



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DOI: https://doi.org/10.22146/jrekpros.24868

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