PMID- 9177706
OWN - NLM
STAT- MEDLINE
DCOM- 19970710
LR  - 20171116
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 247
IP  - 2
DP  - 1997 May 1
TI  - Efficient immobilization of proteins by modification of plate surface with 
      polystyrene derivatives.
PG  - 412-6
AB  - Immobilization of proteins on microplate wells by simple adsorption (e.g., for 
      ELISA) is convenient, but it can be inefficient, especially if proteins are 
      hydrophilic or small in size. This problem was alleviated by the use of 
      polyvinylbenzyl lactonoylamide (PVLA). PVLA is strongly adsorbed to the 
      hydrophobic well surface, and its lactonamide part can be oxidized with periodate 
      to generate aldehydo groups. Proteins are then immobilized covalently to the 
      aldehydo groups by reductive amination under mild conditions. Using this method, 
      henceforth termed the PVLA method, alkaline phosphatase (AP) was immobilized to 
      microplates six- to sevenfold greater than by simple adsorption (as measured by 
      activity). Similarly, the activity of immobilized mannose-binding protein A 
      (MBP-A) was 4- to 8-fold higher by the PVLA method than by simple adsorption. The 
      PVLA-coated plates needed as little as 200 ng of MBP-A per well to have a 
      sufficient amount of MBP-A immobilized for the measurement of binding of 
      125I-labeled mannosylated bovine serum albumin (125I-Man-BSA), but unmodified 
      plates required as much as 20 micrograms/well MBP-A to obtain the same response. 
      Recommended conditions for the PVLA method are 40 microliters of 2 mg/ml of PVLA 
      for coating, 1 mM NaIO4 for the generation of the aldehydo groups, and a 2-h 
      reductive amination at 37 degrees C between pH 8 and 9 for the protein ligation.
FAU - Suzuki, N
AU  - Suzuki N
AD  - Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218, USA.
FAU - Quesenberry, M S
AU  - Quesenberry MS
FAU - Wang, J K
AU  - Wang JK
FAU - Lee, R T
AU  - Lee RT
FAU - Kobayashi, K
AU  - Kobayashi K
FAU - Lee, Y C
AU  - Lee YC
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Amines)
RN  - 0 (Carrier Proteins)
RN  - 0 (Enzymes, Immobilized)
RN  - 0 (Mannose-Binding Lectin)
RN  - 0 (Polystyrenes)
RN  - 0 (Proteins)
RN  - 10450-60-9 (Periodic Acid)
RN  - 143107-68-0 (mannose binding protein A)
RN  - 27432CM55Q (Serum Albumin, Bovine)
RN  - 96910-25-7 (poly-N-4-vinylbenzyllactonamide)
RN  - B45A1BUM4Q (metaperiodate)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
RN  - J2B2A4N98G (Lactose)
SB  - IM
MH  - Adsorption
MH  - Alkaline Phosphatase
MH  - Amines
MH  - Carrier Proteins
MH  - Enzymes, Immobilized
MH  - Lactose/*analogs & derivatives
MH  - *Mannose-Binding Lectin
MH  - Oxidation-Reduction
MH  - Periodic Acid
MH  - *Polystyrenes
MH  - *Proteins
MH  - Serum Albumin, Bovine
MH  - Surface Properties
EDAT- 1997/05/01 00:00
MHDA- 1997/05/01 00:01
CRDT- 1997/05/01 00:00
PHST- 1997/05/01 00:00 [pubmed]
PHST- 1997/05/01 00:01 [medline]
PHST- 1997/05/01 00:00 [entrez]
AID - S0003-2697(97)92094-7 [pii]
AID - 10.1006/abio.1997.2094 [doi]
PST - ppublish
SO  - Anal Biochem. 1997 May 1;247(2):412-6. doi: 10.1006/abio.1997.2094.