View All Items

Title: Evaluation of the specificity and sensitivity of monoclonal antibodies produced against sailfish albumin.
Creator: Rossi, Edmund Anthony., Florida Atlantic University, Hartmann, James X.
Abstract/Description: Monoclonal antibodies (MAbs) were produced against sailfish (Istiophorus platypterus) albumin and investigated for their potential use in species identification. Balb/c mice were immunized with albumin purified from sailfish serum. Hybridomas were screened by enzyme-linked immunosorbent assay (ELISA) for reactivity with the purified albumins of sailfish, blue marlin (Makaira nigricans) and white marlin (Tetrapturus albidus). MAbs from 18 different hybridomas exhibited activity against...
Show moreMonoclonal antibodies (MAbs) were produced against sailfish (Istiophorus platypterus) albumin and investigated for their potential use in species identification. Balb/c mice were immunized with albumin purified from sailfish serum. Hybridomas were screened by enzyme-linked immunosorbent assay (ELISA) for reactivity with the purified albumins of sailfish, blue marlin (Makaira nigricans) and white marlin (Tetrapturus albidus). MAbs from 18 different hybridomas exhibited activity against sailfish albumin. Sixteen MAbs showed cross-reactivity with the marlin species. Two MAbs displayed distinct specificity for sailfish albumin. One of these MAbs (M2D1) was conjugated to horseradish peroxidase to construct an ELISA for the identification of sailfish from serum. The assay correctly identified 18 sailfish from 36 billfish samples. The MAb-peroxidase conjugate was highly specific toward sailfish with no detectable reaction against the heterologous species.
Show less
Date Issued: 1992
PURL: http://purl.flvc.org/fcla/dt/14801
Subject Headings: Sailfish, Monoclonal antibodies
Format: Document (PDF)

Title: ANTI-DNA ANTIBODIES IN SYSTEMIC LUPUS.
Creator: Cavallo, Michelle Fay, Hartmann, James X., Florida Atlantic University, Department of Biological Sciences, Charles E. Schmidt College of Science
Abstract/Description: Two novel methodologies were developed for purification and functional (DNA hydrolytic) assessment of anti-DNA antibodies of IgG isotype from patients with Systemic Lupus Erythematosus (SLE). Earlier protocols for purification and analysis of antibody hydrolytic abilities were lengthy, laborious, and potentially disruptive to antibody function. Purification protocols failed to capture all four IgG subclasses and produced multiple bands outside the range of IgG on electrophoretic separation....
Show moreTwo novel methodologies were developed for purification and functional (DNA hydrolytic) assessment of anti-DNA antibodies of IgG isotype from patients with Systemic Lupus Erythematosus (SLE). Earlier protocols for purification and analysis of antibody hydrolytic abilities were lengthy, laborious, and potentially disruptive to antibody function. Purification protocols failed to capture all four IgG subclasses and produced multiple bands outside the range of IgG on electrophoretic separation. Hydrolysis assays were discontinuous increasing the likelihood of introducing error and making them better suited to analysis of endpoint kinetics rather than reaction kinetics. A two-step, affinity-based purification protocol was developed which utilized magnetic Dynabeads to capture serum components with binding affinity for a thymine 20mer followed by capture of the antibody components of this initial anti-T 20mer serum fraction using Protein G. A fluorescence-based method for real-time, continuous analysis of anti-DNA antibody hydrolytic activity utilizing hydrolysis probes was developed and used to characterize abzyme reaction kinetic parameters. Anti-DNA antibodies demonstrated significantly different Vmax and Km values in the hydrolysis assay (p <0.001) when compared with a DNAse I control.
Show less
Date Issued: 2019
PURL: http://purl.flvc.org/fau/fd/FA00013363
Subject Headings: Systemic lupus erythematosus, Anti-DNA antibodies, DNA antibodies, Antibodies, Catalytic, Autoantibodies--Analysis, Antibodies--isolation & purification
Format: Document (PDF)

Title: Evaluation of monoclonal antibody technology for the species-specific identification of Centropomus undecimalis.
Creator: Potter, Christopher Samuel., Florida Atlantic University, Hartmann, James X.
Abstract/Description: A fusion was conducted using spleen cells from a hyperimmunized Balb/C mouse and a murine myeloma cell line, in an attempt to create monoclonal antibodies specific to parvalbumin from Centropomus undecimalis. The fusion protocol utilized a 24 well culture plate protocol. Initial results indicated the successful generation of antibody producing hybridomas specific to the desired parvalbumin; however, attempts to isolate and clone these cells were unsuccessful. The study results clearly...
Show moreA fusion was conducted using spleen cells from a hyperimmunized Balb/C mouse and a murine myeloma cell line, in an attempt to create monoclonal antibodies specific to parvalbumin from Centropomus undecimalis. The fusion protocol utilized a 24 well culture plate protocol. Initial results indicated the successful generation of antibody producing hybridomas specific to the desired parvalbumin; however, attempts to isolate and clone these cells were unsuccessful. The study results clearly demonstrate some potential drawbacks to using the 24-well fusion protocol versus a 96-well protocol. The 24-well protocol seems to favor rapidly growing non-secreting cells due to the lower dilutions involved. This study was successful in isolating murine polyclonal antiserum reactive in isolating murine polyclonal antiserum reactive with piscine parvalbumin. Further, initial cultures of hybridomas were able to distinguish C. undecimalis from several other species of fishes thus confirming the suitability of parvalbumin as a biomaker protein.
Show less
Date Issued: 1998
PURL: http://purl.flvc.org/fcla/dt/15552
Subject Headings: Monoclonal antibodies, Snook--Identification
Format: Document (PDF)

Title: Cross-reactivity between immunoglobulin G antibodies of whales and dolphins correlates with evolutionary distance.
Creator: Nollens, H. H., Ruiz, C., Walsh, M. T., Gulland, F. M. D., Bossart, Gregory D., Jensen, Eric D., McBain, J. F., Wellehan, J. F. X., Harbor Branch Oceanographic Institute
Date Issued: 2008
PURL: http://purl.flvc.org/fau/fd/FA00007248
Subject Headings: Whales, Dolphins, Immunoglobulin G, Antibodies, Cetacea
Format: Document (PDF)

Title: Detection and purification of lupus B cells.
Creator: Saccocio, Seth., Florida Atlantic University, Hartmann, James X.
Abstract/Description: This study developed a method for positive selection of anti-single stranded (ss)DNA B cells from peripheral blood mononuclear cells (PBMCs) using a twenty nucleotide oligomer of deoxythymidine (oligo(dT) 20mer), coupled to microscopie magnetic beads. Oligo(dT) 20mer specificity for plasma anti-ssDNA antibody (Ab) was established through indirect enzyme linked immunosorbent assay (ELISA) and flow cytometry. A novel method for semi-quantitative detection of ssDNA specific, B cells from PBMC...
Show moreThis study developed a method for positive selection of anti-single stranded (ss)DNA B cells from peripheral blood mononuclear cells (PBMCs) using a twenty nucleotide oligomer of deoxythymidine (oligo(dT) 20mer), coupled to microscopie magnetic beads. Oligo(dT) 20mer specificity for plasma anti-ssDNA antibody (Ab) was established through indirect enzyme linked immunosorbent assay (ELISA) and flow cytometry. A novel method for semi-quantitative detection of ssDNA specific, B cells from PBMC purified peripheral blood that utilized flow cytometric analysis of oligo(dT) 20mer Texas Red labeled cells was developed. Qualitative purification of ssDNA specific, B cells using oligo(dT) coupled magnetic beads was determined through light microscopy and flow cytometric analysis of positive sclected cell populations. Cross-reaetivity of oligo(dT) 20mer with receptors distinct from membrane Ab, resulted in the use of oligo(dC) 20mer as a useful blocking agent. Results show anti-ssDNA Ab titer does not correlate with numbers of peripheral blood ssDNA specific, B cells.
Show less
Date Issued: 2003
PURL: http://purl.flvc.org/fcla/dt/13086
Subject Headings: Systemic lupus erythematosus, B cells, DNA antibodies
Format: Document (PDF)

Title: Rapid, immunologic identification of istiophorid fishes from minute tissue samples.
Creator: Shepard, Scot Robert., Florida Atlantic University, Hartmann, James X.
Abstract/Description: A monoclonal antibody (MAb) produced against sailfish (Istiophorus albicans) serum albumin (SFA)/(Rossi et al. 1992) was used in conjunction with rabbit polyclonal antibodies (PAbs) to formulate a sandwich-style enzyme immunoassay (sEIA). The sEIA specifically identifies sailfish tissues utilizing samples such as heart, kidney, white muscle, red muscle, and blood serum. The assay is sensitive to 20ng of SFA. Of the tissues tested, kidney yields the strongest signal. The entire assay is...
Show moreA monoclonal antibody (MAb) produced against sailfish (Istiophorus albicans) serum albumin (SFA)/(Rossi et al. 1992) was used in conjunction with rabbit polyclonal antibodies (PAbs) to formulate a sandwich-style enzyme immunoassay (sEIA). The sEIA specifically identifies sailfish tissues utilizing samples such as heart, kidney, white muscle, red muscle, and blood serum. The assay is sensitive to 20ng of SFA. Of the tissues tested, kidney yields the strongest signal. The entire assay is carried out in a 3cc syringe and, excluding sample preparation, can be executed in 30 minutes. No specialized equipment or training is required. MAbs were also produced against serum albumin purified from white marlin (Tetrapturus albidus) and Atlantic blue marlin (Makaira nigricans). These Mabs were examined for their utility in identification assays for these species.
Show less
Date Issued: 1994
PURL: http://purl.flvc.org/fcla/dt/15052
Subject Headings: Monoclonal antibodies, Sailfish, Billfishes--Identification, Billfishes
Format: Document (PDF)

Title: Crafting Attractive Non-Covalent Interactions for the Study of β-Hairpins with Long Loops.
Creator: Richaud, Alexis D., Roche, Stéphane P., Florida Atlantic University, Department of Chemistry and Biochemistry, Charles E. Schmidt College of Science
Abstract/Description: In this study, we developed a new peptide motif called β-strap (strap = strand + cap) used to fold β-hairpins of varying length. β-Straps are mean to be short sequences (4 to 8 a-amino acids) forming β-sheets using a judicious combination of non-covalent interactions (NCI) to overcome the entropic penalty inherent to long loop closure. Among those, we proved that a couple of CH-π / NH-π interactions from a tryptophan zipper motif were critical to create a stable packing of the structure. To...
Show moreIn this study, we developed a new peptide motif called β-strap (strap = strand + cap) used to fold β-hairpins of varying length. β-Straps are mean to be short sequences (4 to 8 a-amino acids) forming β-sheets using a judicious combination of non-covalent interactions (NCI) to overcome the entropic penalty inherent to long loop closure. Among those, we proved that a couple of CH-π / NH-π interactions from a tryptophan zipper motif were critical to create a stable packing of the structure. To optimize these interactions, we incorporated unnatural tryptophan derivatives having functionalized indole side chains. Finally, the innate ability of the β-strap to bring β-stand in close contact was exploited to promote macrocyclization of long coiled peptides (up to 16 residues). Then, we studied a more complex β-hairpin loop mimics found at the apex of monoclonal antibodies (mAb) complementary determining region 3 (CDR-H3). Using a set of bioinformatics tools, a search of PDB crystal structures revealed that a large set of mAb crystals possess a β-bulge, located at the edge of CDR-H3 loops. A cluster analysis revealed it has an impressive adaptability towards different H3-loop sizes and conformations. In order to evaluate its function in antibodies, we synthesized several β-hairpin models bearing a prototypical β-bulge. By combining short β-straps and the β-bulge, we were able to design β-hairpin peptides mimic of mAb with a variety of lengths and rigidity while retaining a high degree of folding. Starting from pembrolizumab, the most outstanding blocker of the PD-1/PD-L1 checkpoint currently available in clinic, we scoped ~30 CDR-H3 mAb mimics (H3 loop). As a result, several novel β-hairpin peptide inhibitors of the PD-1/PD-L1 pathway were identified (IC50 <0.3 μM).
Show less
Date Issued: 2023
PURL: http://purl.flvc.org/fau/fd/FA00014154
Subject Headings: Antibodies, Peptides, Biochemistry, β-Hairpins
Format: Document (PDF)

Title: Purification and analysis of autoimmune antibody reactive with single stranded DNA.
Creator: Kats, Anna M., Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: This study evaluated two methods for the isolation and purification of anti-DNA antibodies. A two-step affinity purification with streptavidin (SA) biotinylated oligodeoxythymidine (dT) M-280 and protein G Dynabeadsª was compared to a two step method using Melon(TM) Gel and cellulose DNA. Although Melon gel allowed for faster antibody purification and a higher recovery rate it gave a product of less purity than the magnetic bead method. Further characterization of the antibodies was done by...
Show moreThis study evaluated two methods for the isolation and purification of anti-DNA antibodies. A two-step affinity purification with streptavidin (SA) biotinylated oligodeoxythymidine (dT) M-280 and protein G Dynabeadsª was compared to a two step method using Melon(TM) Gel and cellulose DNA. Although Melon gel allowed for faster antibody purification and a higher recovery rate it gave a product of less purity than the magnetic bead method. Further characterization of the antibodies was done by PhastGel(TM) non-reducing SDS-PAGE and isoelectric focusing in order to analyze purity and confirm the polyclonal nature of anti-DNA antibodies. Agilent 2100, with a higher resolution then SDS-PAGE, revealed possible subclasses of different MW not detected by SDS-PAGE. ELISA showed that all four IgG antibody subclasses were present, while Western blot confirmed the presence of human IgGs. Ultraviolet spectroscopy, Agilent, and fluorescence based assays were used to demonstrate DNA hydrolytic activity of purified anti-DNA antibody.
Show less
Date Issued: 2008
PURL: http://purl.flvc.org/FAU/77646
Subject Headings: DNA antibodies, Monoclonal antibodies, Diagnostic use, Serology, Technique, Systemic lupus erythematosus, Immunological aspects
Format: Document (PDF)

Title: A comparison of several serologic tests to detect antibodies to Toxoplasma gondii in naturally exposed bottlenose dolphins (Tursiops truncatus).
Creator: Dubey, J. P., Fair, Patricia A., Bossart, Gregory D., Hill, D., Fayer, R., Sreekumar, C., Kwok, O. C. H., Thulliez, P., Harbor Branch Oceanographic Institute
Date Issued: 2005
PURL: http://purl.flvc.org/fau/fd/FA00007044
Subject Headings: Bottlenose dolphin, Tursiops truncatus, Toxoplasma gondii, Antibodies, Serologic Tests
Format: Document (PDF)

Title: A study of conjugated monoclonal antibodies in an immunoassay for fish species identification.
Creator: Yu, Wenjie, Florida Atlantic University, Hartmann, James X.
Abstract/Description: Atlantic billfishes (family Istiophoridae) are overexploited and often illegally harvested. To address both of these problems, a rapid means of identifying billfish carcasses is needed. This thesis describes a simple and rapid Nalge Nunc-Immuno(TM) Stick-based direct sandwich assay for sailfish identification that can be performed in the field. A species-specific anti-sailfish monoclonal antibody, covalently bound to the Nalge Nunc-Immuno(TM) Stick's polystyrene surface, was used to capture a...
Show moreAtlantic billfishes (family Istiophoridae) are overexploited and often illegally harvested. To address both of these problems, a rapid means of identifying billfish carcasses is needed. This thesis describes a simple and rapid Nalge Nunc-Immuno(TM) Stick-based direct sandwich assay for sailfish identification that can be performed in the field. A species-specific anti-sailfish monoclonal antibody, covalently bound to the Nalge Nunc-Immuno(TM) Stick's polystyrene surface, was used to capture a biomarker molecule---serum albumin---from sailfish tissue samples. This antibody-antigen interaction was visualized by utilizing peroxidase-conjugated anti-billfish monoclonal antibodies or polyclonal antibodies together with a precipitating substrate. This technique successfully differentiated between sailfish and other fishes within 15 minutes, with a high degree of sensitivity and accuracy. This assay has many potential applications for species identification.
Show less
Date Issued: 2000
PURL: http://purl.flvc.org/fcla/dt/15794
Subject Headings: Monoclonal antibodies, Fishes--Identification, Fishes--Immunology
Format: Document (PDF)

Title: Synthesis and Biological Evaluation of β-hairpin Peptides as Covalent Inhibitors of the PD-1/PD-L1 Immune Checkpoint.
Creator: Naylon, Sarah, Roche, Stephane, Florida Atlantic University, Department of Chemistry and Biochemistry, Charles E. Schmidt College of Science
Abstract/Description: Protein─protein interactions (PPIs) are essential for cell─cell interactions and cellular signal transduction, which play a crucial role in various human diseases. PPIs involved in cancer immunology pathways, known as immune checkpoints, have been intensely studied, leading to a new approach to cancer therapy. The PD1:PDL1 interaction is one of the most essential immune checkpoints. Studies on PD1:PDL1 showed over ten clinical monoclonal antibodies (mAb) used to treat cancer patients....
Show moreProtein─protein interactions (PPIs) are essential for cell─cell interactions and cellular signal transduction, which play a crucial role in various human diseases. PPIs involved in cancer immunology pathways, known as immune checkpoints, have been intensely studied, leading to a new approach to cancer therapy. The PD1:PDL1 interaction is one of the most essential immune checkpoints. Studies on PD1:PDL1 showed over ten clinical monoclonal antibodies (mAb) used to treat cancer patients. Unfortunately, antibodies do not penetrate the tumor microenvironment well, and clearance from the body is slow, leading to unwanted side effects. There is a significant gap in the drug market between the typical Rule of 5 (Ro5) small-molecule drugs (MW<0.5 kDa, SASA ~150 Å) and large antibodies with molecular weights greater than 40 kDa (SASA >2,000 Å). PPIs remain challenging to modulate by small molecules due to their large, shallow, often dynamic, and water-exposed surfaces lacking well-defined binding pockets. Thus, our lab was drawn to work on large β-hairpin peptides (2-3 kDa) that can potentially mimic the CDR-H3 loops of some of the most potent and clinical anti-PD1 antibodies. Exploration of these β-hairpin peptides provided valuable insights into their folding stability, conformational flexibility, passive membrane permeability, and protein-protein interaction (PPI) blocking activities. Additionally, the rational design of TCIs against PD1, specifically targeting a lysine residue, emerged as a strategy to irreversibly obstruct the PD1:PDL1 protein-protein interaction enhancing potency of the non-covalent inhibitors by taking advantage of their specificity. Meticulous structural analysis, peptide synthesis, and biological evaluations are presented contributing comprehensions into covalent inhibitor development of drugs fbRo5.
Show less
Date Issued: 2023
PURL: http://purl.flvc.org/fau/fd/FA00014353
Subject Headings: Cancer--Immunological aspects, Protein-protein interactions, Antibodies, Monoclonal
Format: Document (PDF)

Title: 6-Hydroxydiscodermindole, A New Discodermindole from the Marine Sponge Discodermia polydiscus.
Creator: Cohen, Jesse, Paul, Gopal K., Gunasekera, Sarath P., Longley, Ross E., Pomponi, Shirley A.
Date Issued: 2004
PURL: http://purl.flvc.org/FCLA/DT/2796025
Subject Headings: Antibody-dependent cell cytotoxicity, Calcarea, Demospongiae, Sponges --Caribbean Sea, Sponges --Cytology
Format: Document (PDF)

Title: Systemic Lupus Erythematosus: A.Studies on an ldiotype Specific Dendritic Cell Vaccine. B.Association of Lupus Calcinosis with Calcifying Nanoparticles.
Creator: Keating, Patricia, Florida Atlantic University, Hartmann, James X., Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Systemic lupus erythrematosus (SLE) is an autoimmune disease characterized by the production of anti-DNA antibodies. The primary goal of this study was to activate T cells with specificity toward lupus B cells presenting anti-DNA antibody idiotopes on their surface. Monocyte derived dendritic cells were obtained from peripheral blood of healthy donors and lupus patients. Affinity purified anti-DNA antibodies were obtained from lupus patients' plasmas. The efficacy of different carrier...
Show moreSystemic lupus erythrematosus (SLE) is an autoimmune disease characterized by the production of anti-DNA antibodies. The primary goal of this study was to activate T cells with specificity toward lupus B cells presenting anti-DNA antibody idiotopes on their surface. Monocyte derived dendritic cells were obtained from peripheral blood of healthy donors and lupus patients. Affinity purified anti-DNA antibodies were obtained from lupus patients' plasmas. The efficacy of different carrier proteins, conjugated to lgG, was evaluated, and KLH found to be the most efficient for antigen uptake. The cognate dendritic cells were evaluated for their capacity to activate autologous T cells, and generate a Th1 mediated response which was evaluated by proliferation assays and interferony secretion. During the vaccine study a patient presented with panniculitis, CREST syndrome and a calcinotic exudate. A secondary goal of my study was to analyze this exudate. Calcifying nanoparticles were isolated in lymphocyte culture medium. They were characterized by Von Kassa staining for hydroxyapatite, solubilization by the calcium chelating agent EDT A, and light and scanning electron microscopy. A novel method was developed, using a specific monoclonal antibody, to analyze the calcifying nanoparticles. This method allowed for an approximate quantification of the particles. These particles increased in numbers when incubated for different time periods, and their numbers were decreased when incubated in the presence of minocyclin. Concomitantly, the panniculi in the patient underwent remission with long term antibiotic therapy. CNPs were also obtained from fetal bovine serum and human plasma samples from both lupus patients and healthy donors. Peripheral blood mononuclear cells from healthy donors and lupus patients were analyzed in vitro for their reactivity when incubated in the presence of a biofilm, generated by the calcifying nanoparticles. Viability, proliferation, and co-stimulatory marker up-regulation were determined in the presence or absence of the particles. Osteopontin was found highly expressed in the supernatants of the cells grown with CNPs. Microarray of the mononuclear cells of a healthy donor and a lupus patient incubated in the presence or absence of CNPs was performed, and the results coincided with those determined in vitro.
Show less
Date Issued: 2007
PURL: http://purl.flvc.org/fau/fd/FA00000865
Subject Headings: Systemic lupus erythematosus--Molecular aspects, Systemic lupus erythematosus--Immunological aspects, Cell surface antigens, Autoimmune diseases--Research, Monoclonal antibodies--Diagnostic use
Format: Document (PDF)

FAU Theses and Dissertations (10)	+ -
HBOI Faculty Contributions (3)	+ -

Monoclonal antibodies (5)	+ -
Antibodies (3)	+ -
DNA antibodies (3)	+ -
Systemic lupus erythematosus (3)	+ -
Sailfish (2)	+ -

Anti-DNA antibodies (1)	+ -
Antibodies, Catalytic (1)	+ -
Antibodies, Monoclonal (1)	+ -
Antibodies--isolation & purification (1)	+ -
Antibody-dependent cell cytotoxicity (1)	+ -
Autoantibodies--Analysis (1)	+ -
Autoimmune diseases--Research (1)	+ -
B cells (1)	+ -
Billfishes (1)	+ -
Billfishes--Identification (1)	+ -
Biochemistry (1)	+ -
Bottlenose dolphin (1)	+ -
Calcarea (1)	+ -
Cancer--Immunological aspects (1)	+ -
Cell surface antigens (1)	+ -
Cetacea (1)	+ -
Demospongiae (1)	+ -
Diagnostic use (1)	+ -
Dolphins (1)	+ -
Fishes--Identification (1)	+ -
Fishes--Immunology (1)	+ -
Immunoglobulin G (1)	+ -
Immunological aspects (1)	+ -
Monoclonal antibodies--Diagnostic use (1)	+ -
Peptides (1)	+ -

Electronic Thesis or Dissertation (10)	+ -
article (3)	+ -

Florida Atlantic University (10)	+ -
Hartmann, James X. (7)	+ -
Charles E. Schmidt College of Science (5)	+ -
Department of Biological Sciences (3)	+ -
Bossart, Gregory D. (2)	+ -

Search In

Sort Results By:

Narrow results by:

Collection

Topical Subject

Genre

Type of Resource

Language

Creator

Format

Owner