Produksi dan Karakterisasi Protein Rekombinan L1 HPV 52 Menggunakan Fermentasi Kontinyu dengan Inang Yeast Metilotrofik Hansenula polymorpha

Abstract

Prevalensi kanker serviks secara global menempati urutan keempat terbanyak, salah satunya di Indonesia dengan prevalensi tertinggi disebabkan infeksi Human papillomavirus (HPV) 52. Pengembangan vaksin berbasis protein rekombinan kapsid mayor L1 HPV 52 dengan menggunakan sistem ekspresi yeast melalui metode fermentasi kontinyu belum banyak dilakukan. Penelitian ini bertujuan memproduksi, memurnikan, dan mengkarakterisasi protein rekombinan L1 HPV 52 menggunakan metode fermentasi kontinyu dengan inang yeast Hansenula polymorpha. Proses fermentasi dilakukan dengan bioreaktor 1 L, dilanjutkan dengan lisis sel, purifikasi menggunakan Fast Protein Liquid Chromatography (FPLC), serta karakterisasi melalui Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE), Enzyme-Linked Immunosorbent Assay (ELISA), dan uji protein total bicinchoninic acid (BCA). Hasil SDS-PAGE belum menunjukkan keberadaan pita protein L1, sehingga dikonfirmasi dengan ELISA. Keseluruhan konsentrasi protein L1 HPV 52 diperoleh sebesar 0,0456 µg/mL, dengan recovery yield setelah purifikasi berkisar antara 0,94–5,58%. Rendahnya nilai recovery yield diduga disebabkan oleh agregasi protein serta lemahnya interaksi protein dengan resin kolom.

The prevalence of cervical cancer globally ranks fourth, one of which is in Indonesia with the highest prevalence caused by Human papillomavirus (HPV) 52 infection. The development of a vaccine based on L1 major capsid recombinant protein of HPV 52 using yeast expression system through continuous fermentation method has not been widely done. This study aimed to produce, purify, and characterize L1 HPV 52 recombinant protein using a continuous fermentation method with the yeast host, Hansenula polymorpha. The fermentation process was carried out with a 1 L bioreactor, followed by cell lysis, purification using Fast Protein Liquid Chromatography (FPLC), and characterization through Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), Enzyme-Linked Immunosorbent Assay (ELISA), and total protein bicinchoninic acid (BCA) assay. SDS-PAGE results did not show the presence of L1 protein bands, so it was confirmed by ELISA. The overall concentration of L1 protein HPV 52 obtained was 0.0456 µg/mL, with recovery yield after purification ranging from 0.94-5.58%. The low recovery yield value is thought to be caused by protein aggregation and weak protein interaction with column resin.