Immobilization and recovery of fusion proteins and B-lymphocyte cells using magnetic separation.
A new approach to facilitate immobilization and affinity purification of recombinant proteins and selected human B lymphocytes has been developed. Using magnetic beads with attached DNA containing the Escherichia coli lac operator, fusion proteins comprising the DNA-binding lac repressor could be affinity-purified and recovered by gentle elution conditions, such as with a lactose analogue or by enzymatic means using either deoxyribonuclease (DNase) or restriction endonucleases. The results show for the first time that a DNA-binding protein can be used for affinity purification of fusion proteins as exemplified by the specific and gentle recovery of beta-galactosidase and alkaline phosphatase from bacterial lysates using immunomagnetic separation. The approach was further extended to cell separation by the efficient recovery and elution of human CD37 B lymphocytes from peripheral blood.
Ljungquist
C
C
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Lundeberg
J
J
Rasmussen
A M
AM
Hornes
E
E
Uhlen
M
M
D000469:Alkaline Phosphatase / Q000302:isolation & purification; D000941:Antigens / Q000302:isolation & purification; D001402:B-Lymphocytes / Q000166:cytology*; D001483:Base Sequence; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D008280:Magnetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*; D012097:Repressor Proteins / Q000302:isolation & purification; D013205:Staphylococcal Protein A / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000302:isolation & purification
D000941:Antigens; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D013205:Staphylococcal Protein A; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase