Study on orientation of immunoglobulin G on protein G layer.

A comparative study of immunoglobulin G (IgG) immobilization was performed, both on a thiolated protein G layer, where this immobilization was due to affinity binding with an Fc fragment of IgG, and on 11-mercaptoundecanoic acid (11-MUA), where the immobilization was due to chemical bonding. The change of IgG layer formation on the two base layers as a function of the IgG concentration was investigated by surface plasmon resonance (SPR), atomic force microscopy (AFM) in a non-contact mode, and spectroscopic ellipsometry (SE). It was observed that the IgG layer was immobilized more evenly on the thiolated protein G layer than on the 11-MUA layer, based on the SPR measurements. The surface topology analysis by AFM indicated that the IgG layer was immobilized on the protein G layer according to the envelope profile of the base layer. Based on the SE analysis, it was determined that the IgG layer thickness on the thiolated protein G layer increased with increasing IgG concentration. Based on the above analyses, the scheme for orientation of IgG immobilized on the thiolated protein G layer was proposed.

[1]  M C Davies,et al.  Surface plasmon resonance for real time in situ analysis of protein adsorption to polymer surfaces. , 1997, Biomaterials.

[2]  Se-Hwan Paek,et al.  Strategies for the Immobilization of Antibodies , 1993 .

[3]  Vadim L. Chashchin,et al.  A model of optical immunosensor for hemoproteins based on Langmuir-Blodgett films of FITC-labelled immunoglobulin G , 2000 .

[4]  N. Jaffrezic‐Renault,et al.  Investigating specific antigen/antibody binding with the atomic force microscope. , 2002, Biomolecular engineering.

[5]  J. Halámek,et al.  Highly sensitive detection of cocaine using a piezoelectric immunosensor. , 2002, Biosensors & bioelectronics.

[6]  Martin Hegner,et al.  Ultralarge atomically flat template-stripped Au surfaces for scanning probe microscopy , 1993 .

[7]  G. Whitesides,et al.  Self-assembled organic monolayers: model systems for studying adsorption of proteins at surfaces , 1991, Science.

[8]  David A. Russell,et al.  Self-assembled monolayers: a versatile tool for the formulation of bio-surfaces , 2000 .

[9]  Nikin Patel,et al.  Immobilization of Protein Molecules onto Homogeneous and Mixed Carboxylate-Terminated Self-Assembled Monolayers , 1997 .

[10]  Langmuir films of Fc binding receptors engineered from protein A and protein G as a sublayer for immunoglobulin orientation , 1996 .

[11]  H. Arwin,et al.  Optical Properties of Thin Layers of Bovine Serum Albumin, γ-Globulin, and Hemoglobin* , 1986 .

[12]  A. Ulman,et al.  Formation and Structure of Self-Assembled Monolayers. , 1996, Chemical reviews.

[13]  Hendrik Neubert,et al.  Enhanced affinity capture MALDI-TOF MS: orientation of an immunoglobulin G using recombinant protein G. , 2002, Analytical chemistry.

[14]  T. Lohner,et al.  Comparative study of surface roughness measured on polysilicon using spectroscopic ellipsometry and atomic force microscopy , 1998 .

[15]  Jeong-Woo Choi,et al.  Detection of Escherichia coli O157:H7 using immunosensor based on surface plasmon resonance , 2002 .

[16]  Characterization of IgG Langmuir-Blodgett films immobilized on functionalized polymers. , 2000, Talanta.

[17]  R. Traut,et al.  Addition of sulfhydryl groups to Escherichia coli ribosomes by protein modification with 2-iminothiolane (methyl 4-mercaptobutyrimidate). , 1978, Biochemistry.

[18]  Mandana Veiseh,et al.  Highly Selective Protein Patterning on Gold−Silicon Substrates for Biosensor Applications , 2002 .

[19]  H. Arwin,et al.  Interpretation of Spectroscopic Ellipsometry Data on Protein Layers on Gold Including Substrate-Layer Interactions , 1995 .

[20]  R. Collins,et al.  Correlation of real time spectroellipsometry and atomic force microscopy measurements of surface roughness on amorphous semiconductor thin films , 1996 .

[21]  C. Nicolini,et al.  Comparative Study of Langmuir Monolayers of Immunoglobulin G Formed at the Air-Water Interface and Covalently Immobilized on Solid Supports , 1995 .

[22]  M. Smyth,et al.  Oriented immobilization of antibodies and its applications in immunoassays and immunosensors. , 1996, The Analyst.

[23]  D. De Rossi,et al.  A fluorescent-photochrome method for the quantitative characterization of solid phase antibody orientation. , 2002, Analytical biochemistry.

[24]  M. Aizawa,et al.  Microscopic characterization of Langmuir–Blodgett films incorporating biosynthetically lipid-tagged antibody , 2001 .

[25]  Björn Åkerman,et al.  Design of Oligonucleotide Arrays at Interfaces , 1999 .

[26]  G. Whitesides,et al.  Monolayer Films Prepared by the Spontaneous Self-Assembly of Symmetrical and Unsymmetrical Dialkyl Sulfides from Solution onto Gold Substrates: Structure, Properties, and Reactivity of Constituent Functional Groups , 1988 .

[27]  T. Matsunaga,et al.  Fully automated chemiluminescence immunoassay of insulin using antibody-protein A-bacterial magnetic particle complexes. , 2000, Analytical chemistry.

[28]  Shaoyi Jiang,et al.  Controlling Antibody Orientation on Charged Self-Assembled Monolayers , 2003 .

[29]  B. Chin,et al.  Rapid and sensitive biosensor for Salmonella. , 2000, Biosensors & bioelectronics.

[30]  A. Suleiman,et al.  A Piezoelectric Immunosensor for the Detection of Cocaine , 1996 .

[31]  M. Ibba,et al.  Antibody detection in human serum using a versatile protein chip platform constructed by applying nanoscale self-assembled architectures on gold. , 2002, Journal of proteome research.

[32]  C. Gerber,et al.  Immobilization of Antibodies on a Photoactive Self-Assembled Monolayer on Gold , 1996 .

[33]  Bo Liedberg,et al.  Functionalized Self-Assembled Monolayers on Gold as Binding Matrices for the Screening of Antibody-Antigen Interactions , 2003 .

[34]  C. Meuse.  Infrared Spectroscopic Ellipsometry of Self-Assembled Monolayers , 2000 .