Fabrication of Ceramic Membrane Chromatography for Biologics Purification

Abstract

Chromatography is one of the most important separation processes of choice for the recovery/purification of proteins and complex bio-structures. Fabrication of chromatographic membranes and their efficiency in the chromatography process has been the subject of many recent researches. In this study, a coin-like, 13 mm diameter and 3 mm thick, ceramic membrane was fabricated to be used as a chromatographic medium. The membrane is used to replace the conventional resin-based chromatography columns. Hydroxyapatite (HA) powder was used as a material for the membrane fabrication. In this project, a HA powder was produced using starch as pore creating agents. Characterization processes were done for the ceramic membrane using the suitable apparatuses. Three parameters of the fabrication process (starch wt %, compaction pressure and sintering temperature) were manipulated to optimize the performance of the membrane. The fabricated membrane was placed in a (FPLC) system to be tested for its performance as an adsorptive membrane. (IMAC) process was run by immobilizing Ni2+ ions at the membrane particles surfaces. NP protein of the (NDV) was used to test the membrane's ability to bind Histidine-tagged proteins. The optimum set of process parameters that yielded in the highest porosity and good chromatogram was determined to be 5 wt % starch, 3000 psi compaction pressure and 1100°C sintering temperature.ABSTRAK: Kromatografi merupakan satu daripada proses pengasingan yang penting yang dipilih untuk perolehan/penapisan protein dan biostruktur yang kompleks. Pemfabrikatan membran kromatografi dan kecekapannya dalam proses kromatografi merupakan fokus beberapa kajian terkini. Dalam kajian ini, membran seramik berbentuk duit syiling, berdiameter 13 mm dengan ketebalan 3 mm, direka untuk digunakan sebagai perantara kromatografi. Membran ini digunakan untuk menggantikan turus kromatografi berasaskan resin yang lazim. Serbuk hidroksiapatit (HA) digunakan sebagai bahan pemfabrikatan membran. Dalam projek ini, serbuk HA dihasilkan menggunakan kanji sebagai agen penghasilan liang. Proses pencirian dilakukan terhadap membran seramik menggunakan radas yang sesuai. Tiga parameter proses pemfabrikatan (peratusan berat kanji, tekanan padatan dan suhu pensinteran) dimanipulasikan untuk mendapatkan prestasi membran yang optima. Membran yang difabrikatkan diletakkan dalam sistem FPLC (Fast Protein Liquid Chromatography) untuk diuji prestasinya sebagai membran serap. Proses IMAC (Immobilized Metal Affinity Chromatography) dijalankan dengan memegunkan ion Ni2+ pada permukaan zarah membran. Nucleoprotein dari NDV (Newcastle disease virus) digunakan untuk menguji kebolehan membran terikat dengan protein yang dilabelkan dengan Hisditina. Set parameter proses yang optima yang menghasilkan keliangan tertinggi dan kromatogram yang baik ditentukan pada berat kanji 5 %, tekanan padatan 3000 psi dan suhu pensinteran 1100°C.KEYWORDS: Membrane Chromatography, Porous Ceramic Membrane, IMAC, Hydroxyapatite, Chromatography.

Characterization and Sintering Properties of Hydroxyapatite Bioceramics Synthesized From Clamshell Biowaste

“…The ACS synthesized HA powders were compacted into 20 mm disc samples (Fig. 2) by a hydraulic press machine (Enerpac, USA) at 1000 psi, which was set at a pressure lower than 3000 psi, as recommended by Mel et al [17]. The green samples were then conventionally sintered (Carbolite Gero, UK) at 1200 °C, 1250 °C and 1300 °C for two hours with a ramp rate of 10 °C/min.…”