Date of Award

11-2025

Degree Type

Thesis

Degree Name

MPhil in Biological and Biomedical Sciences

First Advisor

Dr. Fawad Ur Rehman

Second Advisor

Dr. Afsar Ali Mian

Third Advisor

Dr. Irfan Hussain

Department

Biological and Biomedical Sciences

Abstract

Cell isolation is a fundamental technique in biomedical research, with magnetic activated cell sorting (MACS) widely recognized for its high efficiency, ease of use, and non-reliance on trained personnel. However, in Pakistan, access to MACS is restricted by high costs, prolonged import delays, and exchange rate volatility, which emphasizes the need for indigenously developed alternatives. This study aimed to establish an in-house MACS kit by synthesizing and functionalizing iron oxide nanoparticles (IONPs) for the isolation of CD34⁺ hematopoietic stem cells (HSCs). IONPs were prepared via co-precipitation, surface-modified with methoxy-PEG maleimide, and functionalized with anti-CD34 antibodies (antiCD34@mPEGmal@IONPs). Physicochemical characterization confirmed successful nanoparticle synthesis with a narrow size distribution, high colloidal stability, and strong magnetic responsiveness. Simultaneously, PEG coating reduced oxidative stress and improved biocompatibility. Optimization experiments in Kasumi-1 cells identified nanoparticle concentration, incubation period, and magnetic isolation time as important parameters of cell isolation efficiency. Antibody-functionalized IONPs achieved 1.6-fold higher relative isolation efficiency compared to non-functionalized IONPs in a pure culture of HSCs. Additionally, antiCD34@mPEG-mal@IONPs also demonstrated an isolation efficiency of 44.44% compared to commercially available microbeads in peripheral blood mononuclear cells. These findings demonstrate that antibody-functionalized and PEG-coated IONPs have the potential to serve as a cost-effective alternative to commercial MACS kits. Moreover, this study shows promise as a scaffold technology that can be tailored to isolate other cell types. While further refinement is required to improve specificity and yield, this work provides a groundwork for the development of indigenous magnetophoretic cell isolation technologies in Pakistan

First Page

1

Last Page

100

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