Document Detail

Flow injection analysis for simultaneous quantification of prolactin concentration and glycosylation macroheterogeneity in cell culture samples.
MedLine Citation:
PMID:  19003399     Owner:  NLM     Status:  PubMed-not-MEDLINE    
A flow injection analysis (FIA) system is presented for a twostep immunoassay-based determination of the total humanprolactin (hPRL) concentration along with its degree ofglycosylation. Separate measurement of total hPRL and nonglysosylated human prolactin (nG-hPRL) were made using twoflow-through cartridges each containing immobilized antibodiesof different specificity. The antibodies are immobilized on thesurface of a carrier. Glycosylated hPRL (G-hPRL) and, thus, thedegree of glycosylation were calculated by the differencebetween the two specific determinations. Enhanced specificityfor the determination of nG-hPRL was obtained using unfavorablebinding conditions through incorporation of alkaline pH andchaotropic agents into the carrier/dispersion buffer. The assayfor total hPRL and nG-hPRL were each found to be linear withinthe relevant concentration range. The results of the two-stepFIA method were found to agree with those obtained by thestandard methods of ELISA and western blotting while offeringthe advantage of minimal analysis time (10 min) and eliminationof manual manipulations.
M Reinecke; G Stephanopoulos
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Cytotechnology     Volume:  34     ISSN:  0920-9069     ISO Abbreviation:  Cytotechnology     Publication Date:  2000 Nov 
Date Detail:
Created Date:  2008-11-12     Completed Date:  2011-07-14     Revised Date:  2013-02-27    
Medline Journal Info:
Nlm Unique ID:  8807027     Medline TA:  Cytotechnology     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  237-42     Citation Subset:  -    
Department of Chemical Engineering and Biotechnology Process Engineering Center, Massachusetts Institute of Technology, Room 56-469, Cambridge, MA, USA.
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