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      Binding of Fluorescein and Carboxyfluorescein by Normal and Glycosylated Human Serum Proteins


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          The binding of fluorescein and 6-carboxyfluorescein by normal, diabetic and in vitro glycosylated human serum proteins was analyzed by absorption and fluorescence spectroscopy, gel filtration and equilibrium dialysis. The absorption spectra of bound fluorochromes showed red shifts and hypochromic changes, and fluorochrome fluorescence was reduced by serum proteins. Bio-gel P-6 and Sephadex G–200 gel filtration in solvents containing free fluorochrome demonstrated that fluorescein and carboxyfluorescein were bound by serum proteins of varying molecular weights and/or diffusion rates. Equilibrium dialysis at 37°C showed that human sera contained 3.5 ± (SD) 0.4 × 10<sup>––3</sup> Mof fluorescein and carboxyfluorescein binding sites with an average association constant of 3.9 ± 0.2 ×103 M<sup>––1</sup>. Undiluted serum proteins bound 93% of the fluorochrome at a total concentration of 2×10<sup>––4</sup> M or less. Glycosylation of serum proteins in vivo or in vitro did not change the concentration-dependent or pH-dependent binding of fluorescein or carboxyfluorescein.

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          Author and article information

          Ophthalmic Res
          Ophthalmic Research
          S. Karger AG
          04 December 2009
          : 14
          : 6
          : 416-427
          Department of Ophthalmology, Scheie Eye Institute, School of Medicine, University of Pennsylvania, Philadelphia, Pa., USA
          265219 Ophthalmic Res 1982;14:416–427
          © 1982 S. Karger AG, Basel

          Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

          : 10 February 1982
          : 12 May 1982
          Page count
          Pages: 12
          Original Paper

          Vision sciences,Ophthalmology & Optometry,Pathology
          Equilibrium dialysis,Fluorescein,Nonenzymatic glycosylation,Gel filtration,Absorbance red-shift,Carboxyfluorescein,Human serum proteins


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