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Covalent attachment of functionalized cardiolipin on a biosensor gold surface allows repetitive measurements of anticardiolipin antibodies in serum

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Abstract

Antiphospholipid antibodies (aPL) are a relevant serological indicator of antiphospholipid syndrome (APS). A solid-state surface with covalently bound ω-amine-functionalized cardiolipin was established and the binding of β2-glycoprotein I (β2-GPI) was investigated either by use of surface plasmon resonance (SPR) biosensor, by electrically switchable DNA interfaces (switchSENSE) and by scanning tunneling microscopy (STM). STM could clearly visualize the attachment of β2-GPI to the cardiolipin surface. Using the switchSENSE sensor, β2-GPI as specific ligand could be identified by increased hydrodynamic friction. The binding of anti-cardiolipin antibodies (aCL) was detected against the ω-amine-functionalized cardiolipin-modified SPR biosensor (aCL biosensor) using sera from healthy donors, APS patients and syphilis patients. Our results showed that the aCL biosensor is a much more sensitive diagnostic device for APS patients compared to previous methods. The specificity between β2-GPI-dependent autoimmune- and β2-GPI-independent infection-associated types of aPLs was also studied and they can be distinguished by the different binding kinetics and patterns.

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Abbreviations

aCL:

Anticardiolipin antibody

aPL:

Antiphospholipid antibody

APS:

Antiphospholipid syndrome

AUC:

Area under the curve

aβ2-GPI:

Anti-β2-GPI antibodies

BB:

Blocking buffer

BFP:

Biological false positive

FC:

Flow cell

FCS:

Fetal calf serum

HBS:

HEPES-buffered saline

HRP:

Horseradish peroxidase

LA:

Lupus anticoagulant

mAb:

Monoclonal antibody

MW:

Molecular weight

PBS:

Phosphate-buffered saline

PL:

Phospholipid

ROC:

Receiver operating characteristic

RU:

Resonance units

SAM:

Self-assembled monolayer

SD:

Standard deviation

SLE:

Systemic lupus erythematosus

SPR:

Surface plasmon resonance

STM:

Scanning tunneling microscopy

TMB:

3,3′,5,5′-tetramethylbenzidine

VDRL:

Venereal disease research laboratory

WB:

Washing buffer

β2-GPI:

β2-glycoprotein I

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Acknowledgments

The authors thank Anita Schreiegg for excellent technical assistance. This study was supported by the Stiftung Pathobiochemie und Molekulare Diagnostik of the German Society of Clinical Chemistry and Laboratory Medicine and the Australian Research Council, Discovery Project, Grant DP1094497.

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Authors and Affiliations

  1. Institut für Klinische Chemie und Pathobiochemie, Klinikum rechts der Isar der Technischen Universität München, Ismaninger Straße 22, 81675, Munich, Germany

    Alice Schlichtiger, Markus Thaler & Peter B. Luppa

  2. Department Physik E19, Technische Universität München, James-Franck-Str. 1, 85747, Garching, Germany

    Claudia Baier

  3. School of Chemistry, Bio21 Institute, University of Melbourne, Building 102, 30 Flemington Road, Parkville, VIC, 3010, Australia

    Meng-Xin Yin & Andrew B. Holmes

  4. Dynamic Biosensors, c/o Walter Schottky Institut, Technische Universität München, Am Coulombwall 4, 85748, Garching, Germany

    Makiko Maruyama, Ralf Strasser & Ulrich Rant

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  1. Alice Schlichtiger
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  2. Claudia Baier
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  3. Meng-Xin Yin
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Correspondence to Alice Schlichtiger.

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Schlichtiger, A., Baier, C., Yin, MX. et al. Covalent attachment of functionalized cardiolipin on a biosensor gold surface allows repetitive measurements of anticardiolipin antibodies in serum. Anal Bioanal Chem 405, 275–285 (2013). https://doi.org/10.1007/s00216-012-6467-8

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  • DOI: https://doi.org/10.1007/s00216-012-6467-8

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