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"The purification of recombinant proteins is an important stage in biopharmaceutical research. A commonly used technique is immobilized metal affinity chromatography (IMAC). One of the main advantages of this type of chromatography is that the column can easily be regenerated for subsequent purification work. The mechanism of IMAC is based on bonding between metal ions immobilized on a matrix with a specific amino acid. Because of the strong interactions of the electron donor group on the imidazole ring, histidine is often used in the IMAC purification system. Two types of commercial IMAC resin use a nitrilotriacetic acid (NTA) matrix: a nickel-based (Ni-NTA) and cobalt-based (Co-NTA), better known as TALON. This study was aim to investigate the effect of the metal ions Ni2+ and Co2+ to purify recombinant human erythropoietin (rhEPO) expressed in yeast system Pichia pastoris. The results indicated that both Ni-NTA and Co-TALON gave almost the same level of protein purity; however, Ni-NTA has a higher binding affinity than Co-TALON might be due to the higher stability complex of Ni+. The average amount of protein bound by Ni-NTA and Co-TALON was 183.5 and 38.7 μg/mL, respectively."

"Pengaruh Konsentrasi Mg2+ dan Fe2+ dalam Media Kultur terhadap Pembentukan CGF oleh Mikroalga Chlorella pyrenoidosa INK dan Analisis Asam Amino dengan Kromatografi Cair-Spektrofotometri Massa. Chlorella pyrenoidosa (C. pyrenoidosa ) mengandung Chlorella Growth Factor (CGF), yang terdiri dari protein dan polisakarida. CGF terletak di dalam inti sel dan bermanfaat bagi manusia sebagai suplemen makanan, booster imunitas, dan antioksidan. Pembentukan CGF oleh C. pyrenoidosa dipengaruhi oleh komposisi medium. C. pyrenoidosa INK dikultur dalam media basal dimodifikasi (MBM) dengan berbagai konsentrasi Mg2+ (0,5, 1,0, dan 1,5 g/L) dan Fe2+ (3,5×10-4 dan 5,0×10-4 g/L). Percobaan dilakukan menggunakan rancangan acak lengkap dalam botol 2L dengan tiga kali pengulangan. Hasil penelitian menunjukkan bahwa MBM mengandung Mg2+ 1.0 g/L dan Fe2+ 3.5×10-4 g/L menghasilkan kurva pertumbuhan C. pyrenoidosa yang optimal. Analisis kandungan protein dilakukan dengan metode Lowry menggunakan spektrofotometer pada λ=750 nm, menghasilkan 0,0974 mg/mL (ekstrak) dan 6,4097 mg/mL (supernatan). Selanjutnya, analisis kadar glukosa dilakukan dengan metode fenol sulfat (λ=490 nm), hasil yang diperoleh 49,331 ppm (ekstrak) dan 1566,911 ppm (supernatan). Analisis asam amino dalam CGF menggunakan spektrometri massa kromatografi cair (KC-SM) menunjukkan adanya tirosin, prolin, asam glutamat, alanin, valin, triptopan, fenilalanin, metionin, dan leusin-isoleusin.

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Chlorella pyrenoidosa (C. pyrenoidosa) contains Chlorella Growth Factor (CGF), which consists of protein and polysaccharides. CGF is located inside the nucleus of cells and is beneficial to humans as a food supplement, an immunity booster, and an antioxidant. CGF formation of C. pyrenoidosa is influenced by medium composition. C. pyrenoidosa INK was cultured in a modified basal medium (MBM) with various concentrations of Mg2+ (0.5, 1.0, and 1.5 g/L) and Fe2+ (3.5×10-4 and 5.0×10-4 g/L). The experiments were run and analyzed under a completely randomized design using a 2-L bottle with three replications. The results showed that MBM with 1.0 g/L of Mg2+ and 3.5×10-4 g/L of Fe2+ yielded the optimal growth curve for C. pyrenoidosa. Analysis of protein content was carried out using the Lowry method with a spectrophotometer at λ=750 nm, and the obtained results were 0.0974 mg/mL (extract) and 6.4097 mg/ml (supernatant). Furthermore, analysis of glucose content was carried out using the phenol sulfate method (λ = 490 nm), and the obtained results were 49.331 ppm (extract) and 1566.911 ppm (supernatant). Analysis of amino acids in CGF using liquid chromatography-mass spectrometry (LC-MS) indicated the presence of tyrosine, proline, glutamate, alanine, valine, tryptopan, phenylalanine, methionine, and leucine-isoleucine."