Zwitterionic Polydopamine/Protein G Coating for Antibody Immobilization: Toward Suppression of Nonspecific Binding in Immunoassays

2020 ◽  
Vol 3 (6) ◽  
pp. 3631-3639
Author(s):  
Jihyun Byun ◽  
Soojeong Cho ◽  
Jeong Moon ◽  
Hongki Kim ◽  
Hyunju Kang ◽  
...  
Keyword(s):  
Protein G ◽  
Antibody Immobilization ◽  
Nonspecific Binding

scholarly journals Correction to “Zwitterionic Polydopamine/Protein G Coating for Antibody Immobilization: Toward Suppression of Nonspecific Binding in Immunoassays”

2020 ◽  
Vol 3 (8) ◽  
pp. 5454-5454
Author(s):  
Jihyun Byun ◽  
Soojeong Cho ◽  
Jeong Moon ◽  
Hongki Kim ◽  
Hyunju Kang ◽  
...  
Keyword(s):  
Protein G ◽  
Antibody Immobilization ◽  
Nonspecific Binding

scholarly journals Use of Immunomatrix Methods to Improve Protein-Protein Interaction Detection

2003 ◽  
Vol 2003 (5) ◽  
pp. 291-298 ◽  
Author(s):  
M. Walid Qoronfleh ◽  
Ling Ren ◽  
Daryl Emery ◽  
Maria Perr ◽  
Barbara Kaboord
Keyword(s):  
Protein Interactions ◽  
Protein A ◽  
Light Chains ◽  
Protein G ◽  
Antibody Immobilization ◽  
Protein Protein Interactions ◽  
Protein Protein Interaction ◽  
Interaction Detection ◽  
Main Disadvantage ◽  
Key Techniques

Immunoprecipitation (IP) and coimmunoprecipitation (co-IP) are key techniques for studying protein-protein interactions. These methods utilize immobilized protein A or protein G to isolate antibody-bound target antigens. The main disadvantage of traditional immunoprecipitation and coimmunoprecipitation is that the conditions used to elute the precipitated antigen also release the antibody, contaminating the antigen and destroying the antibody support. To overcome these problems, we describe two methods to generate a reusable antibody support by cross-linking the antibody to immobilized protein A or protein G, or by coupling it directly to the resin. Our studies have demonstrated that the immobilization efficiency for the antibody coupling method was similar for several species of antibody. Furthermore, we illustrate that using both methods of antibody immobilization yields IP and co-IP results similar to traditional protocols but eliminate the antibody heavy and light chains contamination.

scholarly journals Novel UV-activated biofunctionalization of up-converting nanocrystals for detection of proteins

2021 ◽  
Author(s):  
Małgorzata Misiak ◽  
Maciej Gawłowski ◽  
Agnieszka Kowalczyk ◽  
Michał Skowicki ◽  
Katarzyna Prorok ◽  
...  
Keyword(s):  
Lower Energy ◽  
Water Solubility ◽  
Protein G ◽  
Light Activation ◽  
Bacterial Protein ◽  
Nonspecific Binding ◽  
Modified Surface ◽  
Initial Coating ◽  
Biological Environment ◽  
Direct Attachment

AbstractLanthanide doped nanocrystals capable to emit higher energy photons under excitation with lower energy radiation are promising for a broad range of applications including biodetection, biosensing, and bioimaging. However, the adaptation of these nanoparticles to the biological environment that requires good water-solubility, stability and ease of further functionalization still remains a challenge. The application of nanoparticles for biodetection or in various assays encountered many difficulties arising mainly from the strong tendency of nanoparticles for aggregation or nonspecific binding. Here we present a new method to obtain soluble and stable in water-based buffers NaYF4:Yb3+Er3+ nanocrystals with modified surface ready for further conjugation with biomolecules. In the presented approach polyvinylpyrrolidone/vinyl alcohol copolymer (PVP/VA) with photo-activatable linker (N-5-Azido-2-nitrobenzoyl group—ANB-NOS) was used for initial coating due to its high non-covalent affinity to nanoparticles surface. Subsequent coating with aminated dextran by ultraviolet light activation of ANB-NOS was carried out. This step has a significant impact on nanocrystals stability in the physiological buffer as well as on protein conjugation. Conjugation of biomolecules was possible by employing another photo-linker—sulfosuccinimidyl 4,4′-azipentanoate (sulfo-SDA). Bacterial Protein G has been selected to create a universal immune-imaging agent due to its ability to bind Fc fragment of most mammalian immunoglobulins. Moreover, the direct attachment of antibodies to nanoparticles was also examined. The activity of biofunctionalized nanocrystals was tested in immunoblot test, that confirmed preserved activity of attached molecules and lack of undesired nonspecific binding or precipitation on the assay membrane. Graphic abstract

Development of a reusable protein G based SPR immunosensor for direct human growth hormone detection in real samples

2015 ◽  
Vol 7 (23) ◽  
pp. 9875-9884 ◽  
Author(s):  
A. Makaraviciute ◽  
A. Ramanavicius ◽  
A. Ramanaviciene
Keyword(s):  
Growth Hormone ◽  
Human Serum ◽  
Human Growth Hormone ◽  
Human Growth ◽  
Serum Analysis ◽  
Protein G ◽  
Antibody Immobilization ◽  
Real Samples ◽  
Spr Immunosensor

The manuscript focuses on investigating some points in employing protein G based antibody immobilization for the development of a reusable SPR immunosensor aimed at human growth hormone detection in real samples. Following our findings a model immunosensor was developed, tested and adapted for human serum analysis.

Nutrition and osteoporosis

2000 ◽  
Vol 57 (3) ◽  
pp. 152-160 ◽  
Author(s):  
Morselli ◽  
Neuenschwander ◽  
Perrelet ◽  
Lippuner
Keyword(s):  
Vitamin A ◽  
Protein G ◽  
Eine Hohe ◽  
Potential Renal Acid Load ◽  
Acid Load

Um zur Prävention der Osteoporose beizutragen, sollte eine Diät idealerweise folgende Kriterien erfüllen: Osteoporosediät ist keine Kalorienreduktionsdiät, da Körpergewicht und Knochenmasse positiv miteinander assoziiert sind. Die Osteoporosediät soll den jeweiligen Calciumbedarf, der unter anderem vom Alter abhängt, voll abdecken. Fasern der Weizenkleie, Phytat und Oxalat aus Bohnen, Spinat und Rhabarber sowie Koffein sollten mit Vorsicht genossen werden, da sie die gastro-intestinale Calciumabsorption beeinträchtigen. Eine chronische nutritiv-bedingte Säurebelastung des Körpers äußert sich in einer gesteigerten Calciurie und kann sich insbesondere in Kombination mit einer geringen Calciumzufuhr ungünstig auf den Knochen auswirken. Ausschlaggebend ist dabei die PRAL (Potential Renal Acid Load) eines Nahrungsmittels, d.h. dessen Potential, während der Verstoffwechselung Säure zu produzieren. Fleischprodukte, aber auch proteinreiche Käsesorten haben eine hohe PRAL, Milch und Joghurt eine niedrige. Früchte und Gemüse (mit Ausnahme von Spargeln) weisen eine negative PRAL auf, wirken also alkalisierend und sind demzufolge günstig für den Knochenmetabolismus. Die Osteoporosediät verbietet eine hohe Kochsalzzufuhr, da diese via pH-Verschiebung ebenfalls zur erhöhten Calciumausscheidung im Urin führt. Eine ausreichende Eiweißzufuhr muß andererseits gewährleistet sein, damit die für die Knochenmatrix-Synthese essenziellen Aminosäuren zur Verfügung stehen. Das Verhältnis Calcium (mg) zu Protein (g) sollte dabei möglichst 20:1 nicht unterschreiten. Wesentliche Bestandteile der Osteoporose-Diät sind außerdem Magnesium sowie die Vitamine B6 (Kollagenbildung), B12 (Osteoblastenaktivität), C (Kollagensynthese), D (intestinale Kalziumabsorption, Osteoblastenaktivität), E (Knochenwachstum) und K (Gammacarboxylierung von Osteokalzin und Matrixproteinen). Eine exzessive Zufuhr von Vitamin A wirkt sich schädlich auf den Knochen aus. Die Spurenelemente Kupfer, Mangan und Zink sind unentbehrlich für das Knochenwachstum. Phytoöstrogene haben in der vom Menschen aufgenommenen Menge kaum Auswirkungen auf den Knochen.

scholarly journals Immunomodulating nano-adaptors potentiate antibody-based cancer immunotherapy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Cheng-Tao Jiang ◽  
Kai-Ge Chen ◽  
An Liu ◽  
Hua Huang ◽  
Ya-Nan Fan ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Immune Responses ◽  
Cancer Immunotherapy ◽  
Tumor Cells ◽  
Noncovalent Interactions ◽  
Killer Cell ◽  
Antibody Immobilization ◽  
Effector Cells ◽  
Nanoparticle Surface

AbstractModulating effector immune cells via monoclonal antibodies (mAbs) and facilitating the co-engagement of T cells and tumor cells via chimeric antigen receptor- T cells or bispecific T cell-engaging antibodies are two typical cancer immunotherapy approaches. We speculated that immobilizing two types of mAbs against effector cells and tumor cells on a single nanoparticle could integrate the functions of these two approaches, as the engineered formulation (immunomodulating nano-adaptor, imNA) could potentially associate with both cells and bridge them together like an ‘adaptor’ while maintaining the immunomodulatory properties of the parental mAbs. However, existing mAbs-immobilization strategies mainly rely on a chemical reaction, a process that is rough and difficult to control. Here, we build up a versatile antibody immobilization platform by conjugating anti-IgG (Fc specific) antibody (αFc) onto the nanoparticle surface (αFc-NP), and confirm that αFc-NP could conveniently and efficiently immobilize two types of mAbs through Fc-specific noncovalent interactions to form imNAs. Finally, we validate the superiority of imNAs over the mixture of parental mAbs in T cell-, natural killer cell- and macrophage-mediated antitumor immune responses in multiple murine tumor models.

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