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Table 2 Comparison of existing technologies for plasma membrane isolation

From: Designer nanoparticle: nanobiotechnology tool for cell biology

Isolation methodologies Advantages Disadvantages
Density Gradient Centrifugation Conventional method that can be used to isolate plasma membrane along with other subcellular compartments
Effective method to isolate lysosomes from tissue or in vivo cell fractions
Simple procedure that can be performed using an ultracentrifuge
Low yield and low purity
Cannot isolate intact membrane layers
Not efficient for isolating cell membrane lipids and for performing functional studies
Cationic silica based isolation Classical method used to isolate cell membrane layers with high purity
Generic method used to isolate cell membrane from tissue, in vitro and in vivo
Formation of matrix by the use of polylysine crosslinker helps in the isolation of membrane layers without any breakage
Low yield
Isolates only available membrane layer i:e only 50 % of cell surface of adherent cells grown on a petri dish
Not robust to perform lipidomics, glycomics and native condition experiments
Cell Surface biotinylation based pull-down assay Generic method that targets cell surface lysine residue
Can be used in combination with magnetic or non-magnetic beads
Can be used in combination to isolate endosomal compartments
Can isolate only available membrane layer i:e only 50 % of cell surface of adherent cells growing on a petri dish
Technology has not been established to isolate cell membrane from cell suspension
Requires detergent in fractionations
Antibody conjugated magnetic nanoparticle based pulldown assay Can be used to pull down proteins after post-fractionation
Can also be used to target selective micro-domains
Can be used in combination with biotin-streptavidin assay
Isolates only available membrane layer (only 50 % surface of adherent cells on a petri dish
Technology has not been established to isolate cell membrane from cell suspension
requires detergent in fractionations
SPMNPs based plasma membrane isolation; A novel strategy that is generic for any kind of cell systems
Method does not involve use of detergent or antibody that affects the nativity of membranes
Method can also use targeted plasma membrane micro-domain by using ligand-tagging
Can be used to isolate protein under native conditions which can hence be used for functional studies
Can isolate cell membrane lipids
Can perform, first of its kind, cell membrane glycosylation
High yield and high purity
Can be used in combination with endosomal isolations
Technology has not be established for tissue cell membrane isolation and in vivo experiments
Isolates only available membrane layer (only 50 % surface of adherent cells on a petri dish
Technology has not been established to isolate cell membrane from cell suspension. Possibility exists to use this technology for cells in suspension culture