Comprehensive study on binding capacity of human immunoglobulin G to Avid AL affinity gel

Shi, Joseph Y.; Goffe, Randal A.

doi:10.1016/0021-9673(94)00073-5

Cited by 6 publications

(2 citation statements)

References 4 publications

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“…Because nonspecific IgG adsorption to a surface such as glass is often described as irreversible especially over extended periods of time, ,,, it calls into question the exact physical meaning of a binding constant for IgG onto plasma-cleaned glass or as-prepared PDMS stamps. However, the Langmuir model offers the simplest explanation for the observed behavior of IgG adsorption and allows comparison with other protein adsorption studies that use the Langmuir model (e.g., Table ). ,− …”

Section: Resultsmentioning

confidence: 99%

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Scanning Force Microscopy and Fluorescence Microscopy of Microcontact Printed Antibodies and Antibody Fragments

LaGraff

Chu-LaGraff

2006

Langmuir

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Unlabeled primary immunoglobulin G (IgG) antibodies and its F(ab')2 and Fc fragments were attached to oxygen-plasma-cleaned glass substrates using either microcontact printing (MCP) or physical adsorption during bath application from dilute solutions. Fluorescently labeled secondary IgGs were then bound to surface-immobilized IgG, and the relative surface coverage was determined by measuring the fluorescence intensity. Results indicated that the surface coverage of IgG increased with increasing protein solution concentration for both MCP and bath-applied IgG and that a greater concentration of IgG was transferred to a glass substrate using MCP than during physisorption during bath applications. Scanning force microscopy (SFM) showed that patterned MCP IgG monolayers were 5 nm in height, indicating that IgG molecules lie flat on the substrate. After incubation with a secondary IgG, the overall line thickness increased to around 15 nm, indicating that the secondary IgG was in a more vertical orientation with respect to the substrate. The surface roughness of these MCP patterned IgG bilayers as measured by SFM was observed to increase with increasing surface coverage. Physisorption of IgG to both unmodified patterned polydimethylsiloxane (PDMS) stamps and plasma-cleaned glass substrates was modeled by Langmuir adsorption kinetics yielding IgG binding constants of K(MCP) = 1.7(2) x 10(7) M(-1) and K(bath) = 7.8(7) x 10(5) M(-1), respectively. MCP experiments involving primary F(ab')2 and Fc fragments incubated in fluorescently labeled fragment-specific secondary IgGs were carried out to test for the function and orientation of IgG. Finally, possible origins of MCP stamping defects such as pits, pull outs, droplets, and reverse protein transfer are discussed.

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Section: Resultsmentioning

confidence: 99%

“…However, the Langmuir model offers the simplest explanation for the observed behavior of IgG adsorption and allows comparison with other protein adsorption studies that use the Langmuir model (e.g., Table 1). 6,[28][29][30][31][32] IV. Microcontact Printing of IgG Fragments.…”

Section: Langmuir Adsorption Modelmentioning

confidence: 99%