A New Sulfo-Siab Synthesis

Leone-Bay, Andrea; Timony, Peter E.

doi:10.1080/00397918808081324

“…It has to be noted that many crosslinkers, which are perfectly suitable for normal crosslinking protocols (reaction with the preformed complex), such as BS3, are not or only weakly active in the new format. In the novel preactivation format only a few crosslinkers have proven to be suitable: Particularly glutaraldehyde (GA) [50,55,56], succinimidyl iodoacetate (SIA) [53], sulfosuccinimidyl iodoacetate (sulfo-SIA), succinimidyl (4-iodoacetyl)aminobenzoate (SIAB) [48] and sulfosuccinimidyl (4-iodoacetyl)aminobenzoate (sulfo-SIAB) [54] can be recommended. All further experiments have been focused on these pre-selected reagents (Figure 3).…”

Section: Resultsmentioning

confidence: 99%

Preactivation Crosslinking—An Efficient Method for the Oriented Immobilization of Antibodies

Schroeder

¹

,

Xuan

²

,

Völzke

³

et al. 2019

MPs

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Crosslinking of proteins for their irreversible immobilization on surfaces is a proven and popular method. However, many protocols lead to random orientation and the formation of undefined or even inactive by-products. Most concepts to obtain a more targeted conjugation or immobilization requires the recombinant modification of at least one binding partner, which is often impractical or prohibitively expensive. Here a novel method is presented, which is based on the chemical preactivation of Protein A or G with selected conventional crosslinkers. In a second step, the antibody is added, which is subsequently crosslinked in the Fc part. This leads to an oriented and covalent immobilization of the immunoglobulin with a very high yield. Protocols for Protein A and Protein G with murine and human IgG are presented. This method may be useful for the preparation of columns for affinity chromatography, immunoprecipitation, antibodies conjugated to magnetic particles, permanent and oriented immobilization of antibodies in biosensor systems, microarrays, microtitration plates or any other system, where the loss of antibodies needs to be avoided, and maximum binding capacity is desired. This method is directly applicable even to antibodies in crude cell culture supernatants, raw sera or protein-stabilized antibody preparations without any purification nor enrichment of the IgG. This new method delivered much higher signals as a traditional method and, hence, seems to be preferable in many applications.

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“…A similar methodology for density measurement as detailed in our previous work Kosuru et al 13 was adapted in the present study. The standard uncertainty was determined to be 8 × 10 −4 g/ cm 3 for the density measurement.…”

Section: Methodsmentioning

confidence: 99%

“…2,6-Dichlorotoluene is one such compound, which is mainly produced by the reaction between para-toluene sulfonyl chloride and chlorine gas at higher temperatures in the presence of a Lewis acid catalyst. 3 2,6-Dichlorotoluene is primarily used in the production of 2,6dichlorobenzoic acid through oxidation in the presence of a metal catalyst and acetic acid as a solvent. In the production of 2,6-dichlorobenzoic acid using 2,6-dichlorotoluene, several byproducts, namely, 2,6-dichlorobenzyl alcohol and 2,6dichlorobenzaldehyde along with unreacted 2,6-dichloroto-luene and acetic acid are obtained, wherein for the separation of these products, thermodynamic properties such as density, viscosity, molar enthalpy of vaporization, and saturation vapor pressure including the critical properties of the compound come into play.…”

Section: Introductionmentioning

confidence: 99%

Measurement and Correlation Studies of Phase Equilibria and Thermophysical Properties of 2,6-Dichlorotoluene

Bharadwaj,

Kumar,

Kommagalla

et al. 2024

J. Chem. Eng. Data

0

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The present study summarizes the thermophysical properties and vapor pressure data of 2,6-dichlorotoluene, an important intermediate in the agrochemical industries for the production of herbicides. A Swietoslawski-type ebulliometer was used for generating the vapor pressure data experimentally in the range of 1.2−33.2 kPa. The vapor pressure data generated are correlated with the Antoine equation, and it is observed that the Antoine equation exhibited minimal deviations. The critical properties were also estimated using the group contribution methods, mainly Joback, Constantinou, Gani, and Nannoolal methods. The normal boiling point obtained using the Antoine equation is much closer to the boiling point obtained using the Constantinou and Gani method. Furthermore, the thermophysical properties like density, refractive index, and viscosity were experimentally evaluated at 95.5 kPa for the temperature ranging from 293.15 to 333.15 K. The density of the liquids was estimated using the Joback, Constantinou, Gani, and Nannoolal methods by employing the Rackett equation. It was observed that Joback showed the least relative average deviation of 1.442%. Mansingh viscometer is used to measure the viscosity and used the Vogel− Tamman−Fulcher equation for correlating the data and, on comparison, was found to be in good agreement. Apart from this, group contribution methods Nannoolal, Orrick, and Erbar methods were used to estimate the viscosity, wherein the Orrick and Erbar method showed the least deviation among the two. Other than the thermophysical properties and vapor pressure data for the pure component, i.e., 2,6-dichlorotoluene, vapor liquid equilibrium data for the binary system acetic acid (1) + 2,6-dichlorotoluene ( 2) is generated at three different subatmospheric pressures, 66.7, 60, and 40 kPa. NRTL model was used to fit the experimental data, and it was found that the experimental data are in good correlation with the predicted data. The binary interaction parameters for the NRTL model were reported, and T−x−y diagrams were also plotted, which show the experimental and predicted data. Thermodynamic consistency tests were conducted for the vapor−liquid equilibrium data at three different pressures, and it was observed that all of them have passed.

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“…In Table 1, a list of crosslinkers is shown, which have been used for the screening. [46][47][48] GA ++ Succinimidyl iodoacetate [49] SIA ++ Succinimidyl (4-iodoacetyl)aminobenzoate [44] SIAB +++ Sulfosuccinimidyl (4-iodoacetyl)aminobenzoate [44,50] Sulfo-SIAB +++…”

Section: Crosslinker Screeningmentioning

confidence: 99%

Preactivation Crosslinking – An Efficient Method for the Oriented Immobilization of Antibodies

Schroeder¹,

Xuan²,

Völzke³

et al. 2019

Preprint

4

0

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Crosslinking of proteins for their irreversible immobilization on surfaces is a proven and popular method. However, many protocols lead to random orientation and the formation of undefined or even inactive by-products. Most concepts to obtain a more targeted conjugation or immobilization requires the recombinant modification of at least one binding partner, which is often impractical or prohibitively expensive. Here a novel method is presented, which is based on the chemical preactivation of Protein A or G with selected conventional crosslinkers. In a second step, the antibody is added, which is subsequently crosslinked in the Fc part. This leads to an oriented and covalent immobilization of the immunoglobulin with a very high yield. Protocols for Protein A and Protein G with murine and human IgG are presented. This method may be useful for the preparation of columns for affinity chromatography, immunoprecipitation, antibodies conjugated to magnetic particles, permanent and oriented immobilization of antibodies in biosensor systems, microarrays, microtitration plates or any other system, where the loss of antibodies needs to be avoided, and maximum binding capacity is desired. This method is directly applicable even to antibodies in crude cell culture supernatants, raw sera or protein-stabilized antibody preparations without any purification nor enrichment of the IgG. This new method delivered much higher signals as a traditional method and, hence, seems to be preferable in many applications.

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A New Sulfo-Siab Synthesis

Cited by 5 publications

References 1 publication

Preactivation Crosslinking—An Efficient Method for the Oriented Immobilization of Antibodies

Preactivation Crosslinking—An Efficient Method for the Oriented Immobilization of Antibodies

Measurement and Correlation Studies of Phase Equilibria and Thermophysical Properties of 2,6-Dichlorotoluene

Preactivation Crosslinking – An Efficient Method for the Oriented Immobilization of Antibodies

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A New Sulfo-Siab Synthesis

Cited by 5 publications

References 1 publication

Preactivation Crosslinking—An Efficient Method for the Oriented Immobilization of Antibodies

Preactivation Crosslinking—An Efficient Method for the Oriented Immobilization of Antibodies

Measurement and Correlation Studies of Phase Equilibria and Thermophysical Properties of 2,6-Dichlorotoluene

Preactivation Crosslinking &ndash; An Efficient Method for the Oriented Immobilization of Antibodies

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Preactivation Crosslinking – An Efficient Method for the Oriented Immobilization of Antibodies