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Authors: Angel Justiz-Vaillant
Summary: This ELISA was used to study the interactions between Staphylococcal protein-A (SpA) and protein-LA (SpLG) with different immunoglobulin preparations of mammalian and avian species.

Proper citation: Angel Justiz-Vaillant 2021. Protein-A_Protein-LG Sandwich ELISA. protocols.io https://dx.doi.org/10.17504/protocols.io.bsu7nezn Copy   

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Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme

Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. Copy of ELISA for quantification of human C5 in serum or plasma.. protocols.io https://dx.doi.org/10.17504/protocols.io.bj76krre Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Dot blot frequently is underutilized or is not frequently used because it cannot assess the molecular weight of macromolecules, but it is simple to perform and is useful when a particular biomolecule needs to be identified. In this protocol dot blot analysis is used for detection of anti-HIV antibody [1], and it is also available for determination or separation of any protein. It can be very useful in the immunodiagnosis of infectious diseases caused by viruses [1] and bacteria [2] and in addition for the determination of specific DNA sequences by using probes [3]. References1. Justiz Vaillant AA, McFarlane-Anderson N, Akpaka PE, Smikle MP, Ramirez N, et al. (2013) Use of Dot Blots Analysis in the Separation of Anti-HIV Antibodies in Animals. J Chromat Separation Techniq 4: 181. doi:10.4172/2157- 7064.10001812. Cardona-Castro N, Gotuzzo E, Rodriguez M, Guerra H. Clinical application of a dot blot test for diagnosis of enteric fever due to Salmonella enterica serovar typhi in patients with typhoid fever from Colombia and Peru.Clin Diagn Lab Immunol. 2000;7(2):312-313. doi:10.1128/cdli.7.2.312-313.20003. Clément G, Benhattar J. A methylation sensitive dot blot assay (MS-DBA) for the quantitative analysis of DNA methylation in clinical samples.J Clin Pathol. 2005;58(2):155-158. doi:10.1136/jcp.2004.021147

Proper citation: Angel Justiz-Vaillant 2020. Dot Blots Analysis in the Separation of Anti-HIV Antibodies.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjm4kk8w Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Enzyme-linked immunosorbent assay (ELISA) for studying the presence of anti-HIV antibodies in layer hens, rats and cats was a reproducible and feseable test. The presence of the anti-HIV antibodies was further confirmed by dot blot analysis.

Proper citation: Angel Justiz-Vaillant 2020. Enzyme-linked immunosorbent assay (ELISA) for detection of anti-HIV antibodies (ELISA) developed in various animal species. . protocols.io https://dx.doi.org/10.17504/protocols.io.bjmzkk76 Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. Monocytes, T and B cells mostly make IL-24. It causes cancer-specific cell death, causes wound healing and protects against bacterial infections and cardiovascular diseases. [1]Reference1. Justiz Vaillant AA, Qurie A. Interleukin. In:StatPearls. Treasure Island (FL): StatPearls Publishing; June 12, 2019.

Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-24 in human serum.. protocols.io https://dx.doi.org/10.17504/protocols.io.bj3skqne Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Protein LG (SpLG) is an immunoglobulin-binding protein that interacts with the Fab and Fc regions of many mammalian immunoglobulins [1].References1.Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175

Proper citation: Angel Justiz-Vaillant 2020. Direct ELISA for investigating the binding of recombinant or chemically-made Protein-LG  to immunoglobulins.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjxvkpn6 Copy   

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Authors: Angel Justiz-Vaillant, Norma McFarlane-Anderson
Group: University of the West Indies, [email protected]

Proper citation: Angel Justiz-Vaillant, Norma McFarlane-Anderson 2020. Universal sandwich enzyme linked immunosorbent assay for investigating protein-AG (SpAG) interactions with immunoglobulins using a SpL-HRP conjugate.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjpxkmpn Copy   

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Authors: Angel Justiz-Vaillant
Group: Carbon
Summary: This ELISA was used to study the interactions between protein LA (SpLA) and protein-LAG (PLAG) with different immunoglobulin preparations from mammalian and avian species.

Proper citation: Angel Justiz-Vaillant 2021. Chimeric Protein-LA and Protein LAG sandwich ELISA . protocols.io https://dx.doi.org/10.17504/protocols.io.bta7nihn Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]

Proper citation: Angel Justiz-Vaillant 2020. Protein isolation from the extract of coelomic fluid of the Lumbricus terrestris.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjsaknae Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: SpLG comprises of 4 Ig-binding domains of SpL and 2 IgG Fc-binding SpG domains [1]. This hybrid molecule was found to bind many intact human Ig molecules and Ig fragments [1]. It proved a powerful tool for the binding, detection and purification of antibodies [2].It was reported that chimeric SpLG was a potent mitogen for mouse splenic B cells, and induced cell differentiation and the production of immunoglobulins. Inhibition experiments demonstrated that the Ig-binding capacity of both SpG and SpL in the chimeric molecule are independent of each other. It was also shown that SpLG selectively absorbed Igs present in the sera of humans, rabbits, mice and rats [1-3]. The preparation of SpLG-HRP by the periodate method is a novel application [4].References1. Justiz-Vaillant AA, Akpaka PE, McFarlane-Anderson N, Smikle MF. Comparison of techniques of detecting immunoglobulin-binding protein reactivity to immunoglobulin produced by different avian and mammalian species.West Indian Med J. 2013;62(1):12-20.2. Kihlberg BM, Sjöbring U, Kastern W, Björck L. Protein LG: a hybrid molecule with unique immunoglobulin binding properties. J Biol Chem 1992, 267(35): 25583-8.3. Justiz-Vaillant AA, McFarlane-Anderson N, and Smikle M. “Bacterial Immunoglobulin (Ig)-Receptors: Past and Present Perspectives.” American Journal of Microbiological Research, vol. 5, no. 2 (2017): 44-50. doi: 10.12691/ajmr-5-2-4.4. Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175.

Proper citation: Angel Justiz-Vaillant 2020. Preparation of a protein-LG conjugated to horseradish peroxidase by the periodate method.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjk3kkyn Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Avian IgY is structurally and genetically different from its mammalian counterpart IgG. This reagent can be used in ELISA, Western blotting and Dot blot to detect specific chicken IgY to infectious agents or immune system proteins [1]. IgY can be isolated by the Polson Method [2] or the water dilution method [3]. Chicken IgY can crossed-react with IgY from diverse bird species [4]. Similarly avian IgY can be isolated from their egg yolk using the same methodology with modifications that was developed for the chicken IgY isolation [5].Reference1. Larsson A, Bålöw RM, Lindahl TL, Forsberg PO. Chicken antibodies: taking advantage of evolution--a review. Poult Sci. 1993;72(10):1807-1812. doi:10.3382/ps.07218072.Polson A (1990) Isolation of IgY from the yolks of eggs by a chloroform polyethylene glycol procedure. Immunol Invest 19: 253-258. 3. Nilsson E, Hanrieder J, Bergquist J, Larsson A. Proteomic characterization of IgY preparations purified with a water dilution method.J Agric Food Chem. 2008;56(24):11638-11642. doi:10.1021/jf802626t4. Vaillant AJ, McFarlane-Andersonv N, Wisdom B, Mohammed W, Vuma S, et al. (2013) Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA). J Anal Bioanal Tech 4: 175. doi:10.4172/2155-9872.1000175 5. : Justiz Vaillant AA, Ramirez N, Cadiz A, Ferrer B, Akpaka P, et al. (2013) Separation and Reactivity of Avian Immunoglobulin Y. J Chromat Separation Techniq 4: 173. doi:10.4172/2157-7064.1000173

Proper citation: Angel Justiz-Vaillant 2020. Preparation of horseradish peroxidase (HRP) conjugated to chicken anti-IgY . protocols.io https://dx.doi.org/10.17504/protocols.io.bjmvkk66 Copy   

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Authors: Angel Justiz-Vaillant, Monica F. Smikle
Group: University of the West Indies, [email protected]

Proper citation: Angel Justiz-Vaillant, Monica F. Smikle 2020. Universal sandwich ELISA for investigating the binding of Protein-AG (SpAG) to avian immunoglobulins using a peroxidase-labeled -anti-IgY conjugate.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjqzkmx6 Copy   

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Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]
Summary: IFN-γ is a key player in driving cellular immunity. It is capable of orchestrating numerous and vital protective functions to enhance immune responses in cancers and infectious diseases. It exhibits its immunomodulatory effects by enhancing antigen processing and presentation,, inducing an anti-viral state, increasing leukocyte trafficking, boosting the anti-microbial properties of immune cells and by affecting cellular proliferation and apoptosis. [1]ReferenceKak G, Raza M, Tiwari BK. Interferon-gamma (IFN-γ): Exploring its implications in infectious diseases.Biomol Concepts. 2018;9(1):64-79. Published 2018 May 30. doi:10.1515/bmc-2018-0007

Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of interferon gamma (IFN-γ) in human serum or plasma.. protocols.io https://dx.doi.org/10.17504/protocols.io.bj68krhw Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: 1. IBP-HRP conjugates can be prepared in the laboratory by the periodate method.2. The Human IgG used can be commercially available of prepared in the laboratory from human sera by affinity chromatography (using an SpG-agarose column).3. SpG-agarose column are widely commercially prepared.4. PURE-1A antibody purification kit will be discontinued. However there are others product for this purpose as Montage Antibody Purification Kit with PROSEP-G media.

Proper citation: Angel Justiz-Vaillant 2020. Binding properties of Human IgG to immunoglobulin-binding proteins tested by double immunodiffusion (Ouchterlony) technique.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjskkncw Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: The peroxidase-labeled anti-chicken IgY conjugate cross-reacts with many IgY present in the egg of many and diverse avian species. References1. Justiz Vaillant AA, Ramirez N, Cadiz A, Ferrer B, Akpaka P, et al. (2013) Separation and Reactivity of Avian Immunoglobulin Y. J Chromat Separation Techniq 4: 173. doi:10.4172/2157-7064.1000173

Proper citation: Angel Justiz-Vaillant 2020. Direct ELISA for investigating the binding of peroxidase-labeled anti-chicken IgY conjugate with avian immunoglobulins. protocols.io https://dx.doi.org/10.17504/protocols.io.bjxykppw Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Chemical synthesis facilitates the generation of peptides which are very difficult to express in bacteria, peptide/protein backbone modification, the incorporation of unnatural amino acids, and the production or synthesis of D-proteins. The C-terminal cysteine can be added to the amino acidic sequences of HIV peptides (fragment 579-601 of the HIV-gp41 and fragments 308-331 or 421-438 of the HIV-gp120). These peptide fragments were dimerized by cysteine oxidation with dimethyl-sulfoxide [1] to faciliutate their conjugation to keyhole limpet hemocyanin that acts as a carrier protein. Reference:1. Tam JP, Wu CR, Liu w, Zhang JW (1991) Disulfide bond formation in peptides by dimethyl sulfoxide. Scope and applications. J Am Chem Soc 113: 66576662.

Proper citation: Angel Justiz-Vaillant 2020. Conjugation of peptide fragment 579-601 of HIV-gp41 or fragments 308-331 or 421-438 of the HIV-gp120 with keyhole limpet hemocyanin (KLH).. protocols.io https://dx.doi.org/10.17504/protocols.io.bjhukj6w Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukin-10 is a pleiotropic cytokine playing a critical role as a regulator of myeloid and lymphoid cell function. Due to the ability of IL-10 to blocking cytokine synthesis and many accessory cell functions of antigen-presenting cells such as macrophages this cytokine is a potent suppressor of the effector functions of macrophages, T-cells and NK cells. IL-10 also participates in regulating proliferation and differentiation of B-lymphocytes, mast cells and thymocytes [1]. Reference1. Ouyang W, O'Garra A. IL-10 Family Cytokines IL-10 and IL-22: from Basic Science to Clinical Translation.Immunity. 2019;50(4):871-891. doi:10.1016/j.immuni.2019.03.020

Proper citation: Angel Justiz-Vaillant 2020. ELISA for quantification of IL-10 in human serum.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjy4kpyw Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]

Proper citation: Angel Justiz-Vaillant 2020. Indirect ELISA for detection of anti-HIV antibodies in cats immunized with a HIV gp120 peptide.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjnpkmdn Copy   

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Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Interleukin17A is a pro-inflammatory cytokine. This protein is produced by a group of T helper cell known as T- helper17 lymphocytes in response to their stimulation with IL-23. After interaction with its receptor, IL-17 activates signalling pathways that stimulate the production of chemokines [1 ].Reference1. Cho ML, Kang JW, Moon YM, Nam HJ, Jhun JY, Heo SB, Jin HT, Min SY, Ju JH, Park KS, Cho YG, Yoon CH, Park SH, Sung YC, Kim HY (May 2006). Journal of Immunology. 176 (9): 5652–61.

Proper citation: Angel Justiz-Vaillant 2020. Enzyme linked immunosorbent assay (ELISA) for determining the serum concentration of IL-17 in humans.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjyykpxw Copy   

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Authors: Angel Justiz-Vaillant, Belkis Ferrer-Cosme
Group: University of the West Indies, [email protected]

Proper citation: Angel Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of human C3 in serum or plasma.. protocols.io https://dx.doi.org/10.17504/protocols.io.bj72krqe Copy   

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