Influence of buffer composition on the distribution of inkjet printed protein molecules and the resulting spot morphology

Mujawar, Liyakat Hamid; Amerongen, A. van

doi:10.1016/j.talanta.2012.06.006

Cited by 14 publications

(19 citation statements)

References 27 publications

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“…6,11,12 It is a big challenge to attain high quality microarrays on non-porous substrates as the spot morphology and the signal to noise ratio depend on parameters such as substrate material and buffer (pH and composition). 13,14 Due to its easy availability and inexpensive nature, glass is an excellent choice as a substrate material. However, on hydrophilic glass, the spots of printed biomolecules are irregular in shape due to minimum initial binding of the biomolecules with the surface.…”

Section: Introductionmentioning

confidence: 99%

“…A preliminary test method was developed, by combining a rapid PCR method with a userfriendly and fast antibody microarray, the nucleic acid microarray immunoassay (NAMIA). 5,14 The double-labeled amplicons were sandwiched between carbon nanoparticle-neutravidin conjugates and immobilized antibodies in a one-step incubation. Sandwiched amplicons were visible as black spots on a white background.…”

Section: Introductionmentioning

confidence: 99%

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Spot morphology of non-contact printed protein molecules on non-porous substrates with a range of hydrophobicities

Mujawar

Amerongen

2013

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Non-contact inkjet printing technology is one of the most promising tools for producing microarrays. The quality of the microarray depends on the type of the substrate used for printing biomolecules. Various porous and non-porous substrates have been used in the past, but due to low production cost and easy availability, non-porous substrates like glass and plastic are preferred over porous substrates. On these non-porous substrates, obtaining spot uniformity and a high signal to noise ratio is a big challenge. In our research work, we have modified pristine glass slides using various silanes to produce a range of hydrophobic glass substrates. The hydrophobicities of the slides expressed in the contact angle (θ) of a sessile drop of water were 49°, 61°, 75°, 88° and 103°. Using a non-contact inkjet printer, microarrays of biotinylated biomolecules (BSA and IgG) were produced on these modified glass substrates, pristine (untreated) glass and also on HTA polystyrene slides. The uniformity of the spots, reflecting the distribution of the biomolecules in the spots, was analyzed and compared using confocal laser scanning microscopy (CLSM). The quality of the spots was superior on the glass slide with a contact angle of ∼75°. We also investigated the influence of the hydrophobicity of the substrate on a two-step, real diagnostic antibody assay. This nucleic acid microarray immunoassay (NAMIA) for the detection of Staphylococcus aureus showed that on highly hydrophilic (θ < 10°) and hydrophobic substrates (θ > 100°) the assay signal was low, whereas an excellent signal was obtained on the substrates with intermediate contact angles, θ ∼ 61° and θ ∼ 75°, respectively.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Spot morphology of non-contact printed protein molecules on non-porous substrates with a range of hydrophobicities

Mujawar

Amerongen

2013

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“…To overcome this challenge, we will address several possible solutions to get uniform surfaces of TFA-treated samples in future studies. From printing biomolecules on surfaces for diagnostic applications it is well-known that an appropriate buffer composition and the addition of a surfactant/detergent to the sample may result in uniform surfaces 36,37 . Changing the hydrophobicity of the Si-plate or the relative humidity upon drying of the samples on the plate may be other ways to solve this issue 38,39 .…”

Section: Resultsmentioning

confidence: 99%

Fourier-transform infrared spectroscopy for monitoring proteolytic reactions using dry-films treated with trifluoroacetic acid

Kristoffersen

Amerongen

Böcker

et al. 2020

Sci Rep

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In this study we explore the potential of using Fourier-transform infrared (FTIR) spectra of trifluoroacetate-protein and peptide complexes for monitoring proteolytic reactions. The idea of treating dry-films of protein hydrolysates with trifluoroacetic acid (TFA) prior to FTIR analysis is based on the unique properties of TFA. By adding a large excess of TFA to protein hydrolysate samples, the possible protonation sites of the proteins and peptides will be saturated. In addition, TFA has a low boiling point when protonated as well as complex-forming abilities. When forming TFA-treated dryfilms of protein hydrolysates, the excess TFA will evaporate and the deprotonated acid (CF 3 coo −) will interact as a counter ion with the positive charges on the sample materials. In the study, spectral changes in TFA-treated dry-films of protein hydrolysates from a pure protein and poultry by-products, were compared to the FTIR fingerprints of untreated dry-films. The results show that time-dependent information related to proteolytic reactions and, consequently, on the characteristics of the protein hydrolysates can be obtained. With additional developments, FTIR on dry-films treated with TFA may be regarded as a potential future tool for the analysis of all types of proteolytic reactions in the laboratory as well as in industry. Fourier-transform infrared (FTIR) spectroscopy is among the well-established methods for the characterisation of proteins and peptides. The repeated amino acid building blocks constructing the backbone of proteins and peptides give rise to multiple distinctive infrared absorption bands (i.e. the amide bands), containing both chemical and structural information. FTIR is therefore frequently used to study and quantify secondary structures of proteins 1-3. At the same time, the protein information in the FTIR spectra has opened the possibility to study a wide range of protein-derived quality features. This includes parameters such as hydration and solvent effects, pH, peptide size distribution and degree of hydrolysis (DH%) 4-9. Currently, peptide size distribution and DH% are measured using laborious and time-consuming techniques. FTIR, on the other hand, represents a rapid alternative which is applicable to an industrial setting. Recent studies have shown that FTIR spectroscopy can be used to predict parameters such as weight-average molecular weight (M w) derived from the peptide size distribution and DH% with high accuracy 10-12. In recent years, enzymatic protein hydrolysis (EPH) has gained significant attention as a versatile processing technology for protein-rich raw materials. For instance, several value-added peptide products have been developed employing EPH on food-processing by-products. In EPH, proteolytic enzymes are used to break down the proteins into smaller fragments, and the resulting protein hydrolysates are very complex, consisting of thousands

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“…The water contact angle (θ) on polystyrene modified paper was 110.9 ± 0.5 , which was higher than reported on a standard HTA polystyrene slide (θ ~92 ). 24,26 Even though Abe et al 25 already demonstrated the application of PS in producing barriers for hydrophilic channels, the consequence of PS modification on the cellulose surface was not reported. A significant difference in advancing and receding contact angle was observed on PS modified filter paper ( Table 1).…”

Section: Goniometer Measurementsmentioning

confidence: 99%

Hexamethyldisilazane Modified Paper as an Ultra-sensitive Platform for Visual Detection of Hg2+, Co2+, Zn2+ and the Application to Semi-quantitative Determination of Hg2+ in Wastewater

2016

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The present article reports the application of hexamethylsilazane (HMDS) modified filter paper for ultrasensitive detection of Hg 2+ , Co 2+ and Zn 2+ . By chemical vapor deposition of HMDS, a highly hydrophilic filter paper was fabricated to a low wetting (hydrophobic) substrate. The water contact angle (θ) of modified paper was ~128 , whereas scanning electron and atomic force microscopy confirmed the surface modification. Using chromogenic reagents, a one-step assay for aforementioned ions was demonstrated onto pristine as well as hydrophobic paper. The assay was completed in less than 10 min and the end-result was in form of a color change that could be easily read by the naked eye. The limit of detection on modified paper was 0.5 ppb, which was 5-order of magnitude superior to that observed on pristine paper. The proposed method was successfully applied for semi-quantitative determination of Hg 2+ ions in real wastewater samples.

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Influence of buffer composition on the distribution of inkjet printed protein molecules and the resulting spot morphology

Cited by 14 publications

References 27 publications

Spot morphology of non-contact printed protein molecules on non-porous substrates with a range of hydrophobicities

Spot morphology of non-contact printed protein molecules on non-porous substrates with a range of hydrophobicities

Fourier-transform infrared spectroscopy for monitoring proteolytic reactions using dry-films treated with trifluoroacetic acid

Hexamethyldisilazane Modified Paper as an Ultra-sensitive Platform for Visual Detection of Hg2+, Co2+, Zn2+ and the Application to Semi-quantitative Determination of Hg2+ in Wastewater

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