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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. Various phagocytes and epithelial cells synthesize Interleukin-34 (IL-34). It enhances IL-6 production and participates in the differentiation and development of antigen-presenting cells including microglia. [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-34 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj36kqre Copy
Authors: Angel Justiz-Vaillant
Group: Carbon
Summary: This ELISA was used to study the interactions between Staphylococcal protein-A (SpA) and protein-AG (SpAG) with different immunoglobulin preparations from mammalian and avian species.
Proper citation: Angel Justiz-Vaillant 2021. Protein A and Protein AG Sandwich ELISA. protocols.io dx.doi.org/10.17504/protocols.io.bsvane2e Copy
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 dx.doi.org/10.17504/protocols.io.bjskkncw Copy
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 dx.doi.org/10.17504/protocols.io.bjmvkk66 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Binding properties of immunoglobulin-binding protein by double immunodiffusion (Ouchterlony) technique. . protocols.io dx.doi.org/10.17504/protocols.io.bjqdkms6 Copy
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 dx.doi.org/10.17504/protocols.io.bj68krhw Copy
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 dx.doi.org/10.17504/protocols.io.bta7nihn Copy
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 dx.doi.org/10.17504/protocols.io.bjk3kkyn Copy
Authors: Angel Justiz-Vaillant, Suzette E. Curtello
Group: University of the West Indies, [email protected]
Summary: Salmonella are motile, flagellated rod-shaped Zoonotic pathogens which may survive with or without oxygen and do not absorb crystal violet stain.They are decolourised by alcohol due to their outer lipopolysaccharide membrane and thin peptidoglycan layer.They belong to the family Enterobacteriaceae and is implicated with typhoid fever and food-borne illnesses.This pathogen is associated with intestinal disease which may become fatal and has negatively impact the health of individuals and various economies globally.The poultry industry is most impacted and vulnerable to the onslaught of this pernicious microbe.The Lipopolysaccharide Somatic O antigen, flagellar H and virulent V antigenic structure determines the serotype designate of Salmonella species [1]. A wide range of antisera is used in the location of the O antigen associated with the carbohydrate constituent of Lipopolysaccharide and H protein antigen linked with the flagella of the bacteria.Salmonellosis is an inflammation of the intestinal mucosal lining by Salmonella bacteria infiltration resulting in painful symptoms.Both man and animal are vulnerable.Salmonellosis is caused by ingestion of Salmonella with food (Wall et al., 1994).The main route of transmission of Salmonella microbe is fecal-oral. Ingestion of small concentration of Salmonella such as 6 cells with contaminated food or water will lead to salmonellosis infection [2].References1. Varma JK, Marcus R, Stenzel SA, et al. Highly resistant Salmonella Newport-MDRAmpC transmitted through the domestic US food supply: a FoodNet case-control study of sporadic Salmonella Newport infections, 2002-2003.J Infect Dis. 2006;194(2):222-230. doi:10.1086/505084.2. Chen W, Martinez G, Mulchandani A. Molecular beacons: a real-time polymerase chain reaction assay for detecting Salmonella.Anal Biochem. 2000;280(1):166-172. doi:10.1006/abio.2000.4518
Proper citation: Angel Justiz-Vaillant, Suzette E. Curtello 2020. Salmonella Typhimurium isolation from specimens. protocols.io dx.doi.org/10.17504/protocols.io.bjjdkki6 Copy
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 dx.doi.org/10.17504/protocols.io.bjxykppw Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Chicken immunization with peptides is inefective if only just the peptides are being inoculated. However, to make the immune response effective the fragment 254-274 of HIV-1 was conjugated with a carrier protein (KLH) that produced a critical immune response, assessed by ELISA, Immunoblot analysis and dot blot [1-4]. The Polson method (1990) can be used effectively to separate the IgY antibody from the egg yolk of immunized chickens [5].References1. Schade R, Pfister C, Italatsch R, Henklien P (1991) Polyclonal antibodies from chicken egg yolk-an alternative to the production of mammalian IgG type antibodies in rabbits. ATLA 19: 403-419. 2. Justiz Vaillant AA, Akpaka PE, Smikle M, McFarlane-Anderson N (2012) In vitro Inhibition of Staphylococcus aureus Isolates by Anti-Anti-Idiotypic Antibodies to Staphylococcal Protein (SpA). J Vaccines Vaccin 3: 127-131.3. Collins AM, Roberton DM, Hosking CS, Flannery GR (1991) Oral immunization with xenogeneic antibodies stimulates the production of systemic and mucosal anti-idiotypic antibodies. Immunology 73: 388-393. 4. Ho DD, Kaplan JC, Rackauskas IE, Gurney ME (1988) Second conserved domain of gp120 is important for HIV infectivity and antibody neutralization. Science 239: 1021-1023. 5. Polson A (1990) Isolation of IgY from the yolks of eggs by a chloroform polyethylene glycol procedure. Immunol Invest 19: 253-258.
Proper citation: Angel Justiz-Vaillant 2020. Chicken immunization with Keyhole limpet hemocynin (KLH)-gp120 fragment (254-274) conjugate raises anti-KLH antibodies in egg yolks.. protocols.io dx.doi.org/10.17504/protocols.io.bjvtkn6n Copy
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 dx.doi.org/10.17504/protocols.io.bjy4kpyw Copy
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 dx.doi.org/10.17504/protocols.io.bjnpkmdn Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Proper citation: Angel Justiz-Vaillant 2020. Universal sandwich ELISA for investigating the binding of avian and mammalian immunoglobulins to Streptococcal protein-G (SpG) using a peroxidase-labeled Protein LAG conjugate (SpLAG-HRP).. protocols.io dx.doi.org/10.17504/protocols.io.bjsfknbn Copy
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 L to avian immunoglobulins using anti-IgY-peroxidase as conjugate.. protocols.io dx.doi.org/10.17504/protocols.io.bjqykmxw Copy
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-LA (SpLA) interactions with immunoglobulins using a SpG-HRP conjugate.. protocols.io dx.doi.org/10.17504/protocols.io.bjpzkmp6 Copy
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. IL-32 is a pro-inflammatory molecule. Natural killer cells and monocytes mainly produce it. IL-32 induces the synthesis of various cytokines including IL-6, and IL-1beta. It inhibits IL-15 production. [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-32 in human serum.. protocols.io dx.doi.org/10.17504/protocols.io.bj35kqq6 Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: This reagent can be used in ELISA, Western blotting and Dot blot to detect antigens and antibodies. It is important in the immunodiagnosis of infectious diseases and other problems. Protein L binds to kappa light chains of immunoglobulins from many animal species including human, mouse, rat, chicken, hamster and pig [1].1. 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.
Proper citation: Angel Justiz-Vaillant 2020. Preparation of horseradish peroxidase (HRP) conjugated Peptostreptococcal protein-L by the periodate method.. protocols.io dx.doi.org/10.17504/protocols.io.bj5ukq6w Copy
Authors: Angel Justiz-Vaillant
Group: University of the West Indies, [email protected]
Summary: Protein LA, a novel hybrid protein, structurally contains 4 of the Ig Fc-binding and 4 of the Ig Fab binding regions on SpA with 4 of kappa light chain-binding sites of protein L. It has a MW of 65 kDa. Protein LA combines the binding properties of the both SpL and SpA and in some cases, give higher binding affinity than either protein alone. The binding of an Ig to SpL does not interfere with binding of another Ig molecule to the SpA domains and vice versa. Protein LA has been shown to bind effectively to immunoglobulins and their fragments from many species of animals [1]. Reference1. 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.
Proper citation: Angel Justiz-Vaillant 2020. Preparation of a protein-LA conjugated to horseradish peroxidase by the periodate method.. protocols.io dx.doi.org/10.17504/protocols.io.bjkzkkx6 Copy
Authors: Angel Justiz-Vaillant
Group: Carbon
Summary: This ELISA was used to study the interactions between Peptostreptococcal protein-L (SpL) and protein LAG (PLAG) with different immunoglobulin preparations from mammalian and avian species.
Proper citation: Angel Justiz-Vaillant 2021. Peptostreptococcal protein L and a Chimeric Protein-LAG sandwich ELISA. protocols.io dx.doi.org/10.17504/protocols.io.bta6nihe Copy
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