Original ArticleCloning and Expression of Recombinant Human GMCSF From Pichia pastoris GS115-A Progressive Strategy for Economic ProductionSrinivasa Babu, K. MTech, PhD; Pulicherla, Krishna Kanth MTech, PhD; Antony, Aju MS; Meenakshisundaram, Sankaranarayanan MTech, PhDAuthor Information 1Centre for Biotechnology, Anna University, Chennai, India; and 2Centre for Bioseparation Technology, VIT University, Vellore, India. Address for correspondence: Centre for Bioseparation Technology, VIT University, Vellore 632014, India. E-mail: [email protected] The authors have no conflicts of interest to declare. American Journal of Therapeutics: November/December 2014 - Volume 21 - Issue 6 - p 462-469 doi: 10.1097/MJT.0000000000000040 Buy Metrics Abstract Human granulocyte–macrophage colony-stimulating factor (hGMCSF) is a proinflammatory cytokine and hematopoietic growth factor. Recombinant human granulocyte–macrophage colony-stimulating factor (rhGMCSF) serves as a biotherapeutic agent in bone marrow stimulations, vaccine development, gene therapy approaches, and stem cell mobilization. The objective of the present study includes construction of rhGMCSF having N-terminal intein tag, expression of protein both extracellularly and intracellularly from yeast expression system followed by its purification in a single step by affinity chromatography. The soluble and biologically active rhGMCSF was obtained from Pichia pastoris GS115. About 122 g DCW/L of final yield was obtained for both cytosolic and secretory expression of Pichia GS115 strain. Purified intracellular hGMCSF was 420 mg/L with a specific activity of 2.1 × 108 IU/mg, and the purified extracellular recombinant protein was 360 mg/L with a specific activity of 1.9 × 108 IU/mg. The data presented here indicate the possibilities of exploring the economic ways of producing the rhGMCSF. Copyright © 2014 Wolters Kluwer Health, Inc. All rights reserved.