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Adsorption of IgG on/in a PAH/PSS Multilayer Film: Layer Structure and Cell Response
KTH, School of Chemical Science and Engineering (CHE), Chemistry, Surface and Corrosion Science.
KTH, School of Chemical Science and Engineering (CHE), Chemistry, Surface and Corrosion Science.
Mabtech AB, Sweden.
KTH, School of Chemical Science and Engineering (CHE), Chemistry, Surface and Corrosion Science. Institute for Surface Chemistry, Sweden.ORCID iD: 0000-0001-7496-1101
2011 (English)In: Journal of Colloid and Interface Science, ISSN 0021-9797, E-ISSN 1095-7103, Vol. 354, no 1, 31-37 p.Article in journal (Refereed) Published
Abstract [en]

The binding of immunogloblulins (IgG) (mouse monoclonal recognizing IFN gamma) on precoated polystyrene or silica surfaces by the layer-by-layer technique has been investigated with QCM-D and DPI. The aim of the work was to increase the sensitivity of the conventional enzyme-linked immunosorbent spot (ELISpot) assay. The polyelectrolytes used to build the multilayers were poly(allylamine hydrochloride) (PAH)/poly(sodium 4-styrenesulfonate) (PSS) alternately adsorbed from 150 mM NaCl. The multilayer build up is linear and the internal structure of the PAH/PSS multilayer is compact and rigid as observed by low relative water content (20-25%) and high layer refractive index (n similar to 1.5) after the formation of five bilayers. Incorporation of IgG within the PAH/PSS multilayer did not give rise to overcharging and did not affect the linear build up. ELISpot test on PAH/PSS multilayer modified polystyrene wells showed that the cytokine response was significantly smaller than on the regular PVDF backed polystyrene wells. This may be due to the compact and rigid nature of the PAH/PSS multilayer, which does not allow formation of the kind of three dimensional support needed to achieve bioactive IgG binding to the surface. Immunological tests of the polyelectrolyte multilayers in the absence of IgG showed that PSS terminated PAH/PSS multilayer did not induce any cytokine response whereas PAH terminated did, which suggests that PSS totally covers the surface from the cells point of view.

Place, publisher, year, edition, pages
Elsevier , 2011. Vol. 354, no 1, 31-37 p.
Keyword [en]
PAH, PSS, Polyelectrolyte multilayer, Layer-by-layer deposition, IgG, Incorporation, Adsorption, QCM, DPI, ELISpot, Cytokine, Cell response
National Category
Physical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-13938DOI: 10.1016/j.jcis.2010.10.006ISI: 000286121700005Scopus ID: 2-s2.0-78650171483OAI: oai:DiVA.org:kth-13938DiVA: diva2:328466
Funder
Swedish Research Council
Note

QC 20100705

Updated from submitted to published 20110317

Available from: 2010-07-05 Created: 2010-07-05 Last updated: 2016-04-29Bibliographically approved
In thesis
1. Structures of Polyelectrolyte Multilayers and Preasorbed Mucin: The Influence of Counterions
Open this publication in new window or tab >>Structures of Polyelectrolyte Multilayers and Preasorbed Mucin: The Influence of Counterions
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The focus in this thesis has been to gain a fundamental understanding of how different type of salts affect preadsorbed polyelectrolytes, both natural and synthetic. The knowledge from the fundamental work is then applied on a commercial system to investigate if the efficiency can be enhanced.

We built thin films using the synthetic polyelctrolytes by using layer-by layer (LbL) deposition. The formed film is commonly known as a polyelectrolyte multilayer. The LbL method allows the incorporation of proteins, polymers, polyelectrolytes with different functions and so on within the film, thus achieving multilayers with different functions.

The major measuring technique used within this thesis is the quartz crystal microbalance with dissipation (QCM-D), which measures mass adsorbed on a surface including the trapped solvent and the viscoelastic properties of an adsorbed film. The QCM-D measurements were complemented with an optical technique, dual polarization interferometry (DPI), which measures the change in refractive index and thickness. From these parameters the dry mass and relative water content of the film can be calculated. The Atomic Force Microscopy (AFM) further gave information about forces acting between preadsorbed films.

We investigated the effect of salt on synthetic polyelectrolyte poly(allylamine hydrochloride)/poly(sodium 4-styrenesulfonate) built with the LbL technique, thus forming polyelectrolyte multilayers. We concluded that the multilayer build-up was linear and that the internal structure of the multilayer is of a compact and rigid nature. However, the type of rinsing protocol (termination of adsorption by: salt, water and salt first followed by water) has a significant effect on the outer layer of the formed multilayer. Interestingly, the structural changes only applied when poly(allylamine hydrochloride) was at the outermost layer and the most significant when water was used. We suggest that it is only the top layer that swells due to the removal of counterions resulting in increased intrachain repulsion. We further performed two-layer model calculations with the Voight model to confirm the QCM-D results as well as a novel two layer model simulation for the DPI data in order to resolve the thickness. The model calculations were in good agreement with each other thus we concluded that only the outer layer swells for this particular multilayer system.

In a related experiment we studied the adsorption of bovine submaxillary mucin (BSM), which has an important mucousal function, to different thiol modified gold surfaces as well as the effect of electrolytes (NaCl, CaCl2, LaCl3) on preadsorbed mucin to a hydrophobic thiol-modified Au surface. The salt induced an expansion at low concentrations; higher concentrations resulted in a compaction. Increasing the valence of the counter ion resultedin a compaction at low concentrations. The structural change of preadsorbed BSM was reversible for NaCl, partially reversible for CaCl2 and irreversible for LaCl3. Interestingly, the swelling of BSM could not be fully understood by using the QCM-D and thus AFM force curves of the same system were taken and the results showed that NaCl does decrease the tail length due to the effective screening of charged sites within the BSM molecule. Increasing the valence resulted in a notable compaction already at very low concentrations suggesting that the ions bind to the anionic sites on BSM.

In the last work we attempted to combine the gained knowledge from the previous studies by using the LbL-buildup on an actual commercial health care application. The above-mentioned mutlilayer were used to coat polystyrene wells in order to increase the binding of immunoglobulin (IgG). The main goal was to increase the sensitivity of the conventional enzymelinked immunosorbent spot assay (ELISpot) and subsequently the modified polystyrene wells were used with the ELISpot test with human peripheral blood mononuclear cells (PBMC) to measure the cytokine response. We suggested that the main driving force for adsorption for IgG on a PAH terminated multilayer is electrostatic attraction, whereas on PSS terminated multilayer the driving force is hydrophobic. Further, we suggested that IgG  does not overcharge the surface and the linearity of the multilayer build-up is not altered when IgG is incorporated within the multilayer structure. We concluded that the cytokine response (spots) on the built multilayers regardless thickness or adsorbed IgG is significantly less than the regular polyvinyldiene fluoride (PVDF) backed ELISpot wells. We suggested that due to the compact and rigid nature of the PAH/PSS multilayer structure it is unable to form the kind of three-dimensional antibody-binding support found in the PVDF membrane. PSS terminated PAH/PSS multilayer did not induce any cytokine response whereas PAH terminated did, which suggests that PSS totally covers the surface from the cells point of view.

 

Place, publisher, year, edition, pages
Stockholm: KTH, 2010. ix, 51 p.
Series
Trita-CHE-Report, ISSN 1654-1081 ; 2010:18
National Category
Physical Chemistry
Identifiers
urn:nbn:se:kth:diva-13182 (URN)978-91-7415-639-3 (ISBN)
Public defence
2010-06-11, E1, Lindstedtsvägen 3, entréplan, KTH, Stockholm, 10:00 (English)
Opponent
Supervisors
Note
QC20100705Available from: 2010-06-01 Created: 2010-06-01 Last updated: 2010-07-05Bibliographically approved

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