Antibody-conjugated paramagnetic nanobeads: kinetics of bead-cell binding.
Specific labelling of target cell surfaces using antibody-conjugated paramagnetic nanobeads is essential for efficient magnetic cell separation. However, studies examining parameters determining the kinetics of bead-cell binding are scarce. The present study determines the binding rates for specific and unspecific binding of 150 nm paramagnetic nanobeads to highly purified target and non-target cells. Beads bound to cells were enumerated spectrophotometrically. Results show that the initial bead-cell binding rate and saturation levels depend on initial bead concentration and fit curves of the form A(1 - exp(-kt)). Unspecific binding within conventional experimental time-spans (up to 60 min) was not detectable photometrically. For CD3-positive cells, the probability of specific binding was found to be around 80 times larger than that of unspecific binding.
Department of Pathobiology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand. swaseem92@yahoo.com.
Shahid
S
Waseem
Department of Physics, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand. maa5652@gmail.com.
Michael A
MA
Allen
Institute for Innovative Learning, Mahidol University, 999 Phuttamonthon 4 Road, Salaya, Nakorn Pathom 73170, Thailand. stefan-schreier@web.de.
Stefan
S
Schreier
Department of Pathobiology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand. rachanee.udo@mahidol.ac.th.
Rachanee
R
Udomsangpetch
Department of Pathobiology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand. sbhakdi@gmail.com.
Sebastian C
SC
Bhakdi
D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000276:immunology / Q000378:metabolism; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D013053:Spectrophotometry
D000906:Antibodies; D017252:CD3 Complex; D018950:Lipopolysaccharide Receptors; D058185:Magnetite Nanoparticles