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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