Preparation and properties of recombinant Clostridium ramosum IgA proteinase. Isolation of Fc-SC and Fab fragments of human secretory IgA

2021 ◽  
pp. 105891
Author(s):  
Michal Krupka ◽  
Leona Raskova Kafkova ◽  
Lucia Barkocziova ◽  
Kristyna Sloupenska ◽  
Diana Brokesova ◽  
...  
Keyword(s):  
Secretory Iga ◽  
Fab Fragments

Large-cluster and combined fluorescent and gold immunoprobes

1996 ◽  
Vol 54 ◽  
pp. 892-893
Author(s):  
Richard D. Powell ◽  
James F. Hainfeld ◽  
Carol M. R. Halsey ◽  
David L. Spector ◽  
Shelley Kaurin ◽  
...  
Keyword(s):  
Metal Cluster ◽  
Sulfonyl Chloride ◽  
Gel Filtration ◽  
Cross Linking ◽  
Site Specific ◽  
Fab Fragments ◽  
Cluster Label ◽  
Covalently Linked ◽  
Texas Red ◽  
Fold Excess

Two new types of covalently linked, site-specific immunoprobes have been prepared using metal cluster labels, and used to stain components of cells. Combined fluorescein and 1.4 nm “Nanogold” labels were prepared by using the fluorescein-conjugated tris (aryl) phosphine ligand and the amino-substituted ligand in the synthesis of the Nanogold cluster. This cluster label was activated by reaction with a 60-fold excess of (sulfo-Succinimidyl-4-N-maleiniido-cyclohexane-l-carboxylate (sulfo-SMCC) at pH 7.5, separated from excess cross-linking reagent by gel filtration, and mixed in ten-fold excess with Goat Fab’ fragments against mouse IgG (obtained by reduction of F(ab’)2 fragments with 50 mM mercaptoethylamine hydrochloride). Labeled Fab’ fragments were isolated by gel filtration HPLC (Superose-12, Pharmacia). A combined Nanogold and Texas Red label was also prepared, using a Nanogold cluster derivatized with both and its protected analog: the cluster was reacted with an eight-fold excess of Texas Red sulfonyl chloride at pH 9.0, separated from excess Texas Red by gel filtration, then deprotected with HC1 in methanol to yield the amino-substituted label.

scholarly journals Intratracheal inoculation of AHc vaccine induces protection against aerosolized botulinum neurotoxin A challenge in mice

npj Vaccines ◽  
2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Changjiao Gan ◽  
Wenbo Luo ◽  
Yunzhou Yu ◽  
Zhouguang Jiao ◽  
Sha Li ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Clostridium Botulinum ◽  
Freeze Drying ◽  
Botulinum Neurotoxin ◽  
Secretory Iga ◽  
Immune Protection ◽  
Dry Powder ◽  
Botulinum Neurotoxin A ◽  
Spray Freeze Drying

AbstractBotulinum neurotoxin (BoNT), produced by Clostridium botulinum, is generally known to be the most poisonous of all biological toxins. In this study, we evaluate the protection conferred by intratracheal (i.t.) inoculation immunization with recombinant Hc subunit (AHc) vaccines against aerosolized BoNT/A intoxication. Three AHc vaccine formulations, i.e., conventional liquid, dry powder produced by spray freeze drying, and AHc dry powder reconstituted in water are prepared, and mice are immunized via i.t. inoculation or subcutaneous (s.c.) injection. Compared with s.c.-AHc-immunized mice, i.t.-AHc-immunized mice exhibit a slightly stronger protection against a challenge with 30,000× LD50 aerosolized BoNT/A. Of note, only i.t.-AHc induces a significantly higher level of toxin-neutralizing mucosal secretory IgA (SIgA) production in the bronchoalveolar lavage of mice. In conclusion, our study demonstrates that the immune protection conferred by the three formulations of AHc is comparable, while i.t. immunization of AHc is superior to s.c. immunization against aerosolized BoNT/A intoxication.

Immunoglobulins and Blood Vessels in the Tonsillar Crypt Epithelium

1973 ◽  
Vol 82 (3) ◽  
pp. 359-369 ◽  
Author(s):  
John F. Schmedtje ◽  
Ann F. Batts
Keyword(s):  
Epithelial Cells ◽  
Blood Vessels ◽  
Plasma Cells ◽  
Border Zone ◽  
Secretory Iga ◽  
Antigenic Determinants ◽  
Crypt Epithelium ◽  
Lower Border

The localization of IgA, IgG, IgM, SP and the relationships of plasma cells and lymphocytes to blood vessels in the tonsillar crypt epithelium were investigated. Immunofluorescent techniques were used that included antisera specific for the two antigenic determinants of external secretory IgA, namely, 4s SP and 7s IgA, and also antisera specific for 7s IgG and 19s IgM. The secretory piece was absent in the crypt epithelium and in most of the crypt lumen. Aggregations of plasmacyte series cells, containing either IgG, IgA, or IgM were present in the crypt epithelium. Mature plasma cells of these aggregations abutted against the walls of blood sinusoids located in the epithelium, which suggested secretion into these sinusoids. All three immunoglobulins were also identified between epithelial cells and small lymphocytes. Postcapillary venules with emigrating small lymphocytes abounded in sub-epithelial sites, and were present at the lower border zone of the epithelium. Lymphocytes in shapes of diapedesis were observed in the endothelium of epithelial blood sinusoids. These observations are in accord with the hypothesis that a “circulation” of many lymphocytes occurs in the epithelium facilitating the activation of any one genetically committed lymphocyte.

scholarly journals Intravenous Vipera berus Venom-Specific Fab Fragments and Intramuscular Vipera ammodytes Venom-Specific F(ab’)2 Fragments in Vipera ammodytes-Envenomed Patients

Toxins ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 279
Author(s):  
Tihana Kurtović ◽  
Svjetlana Karabuva ◽  
Damjan Grenc ◽  
Mojca Dobaja Borak ◽  
Igor Križaj ◽  
...  
Keyword(s):  
Serum Concentration ◽  
Clinical Signs ◽  
Length Of Hospital Stay ◽  
Vipera Berus ◽  
Fab Fragments ◽  
Elimination Half ◽  
Apparent Total Body Clearance ◽  
Total Body ◽  
A Prospective Study ◽  
Vipera Ammodytes

Vipera ammodytes (V. ammodytes) is the most venomous European viper. The aim of this study was to compare the clinical efficacy and pharmacokinetic values of intravenous Vipera berus venom-specific (paraspecific) Fab fragments (ViperaTAb) and intramuscular V. ammodytes venom-specific F(ab’)2 fragments (European viper venom antiserum, also called “Zagreb” antivenom) in V.ammodytes-envenomed patients. This was a prospective study of V.ammodytes-envenomed patients that were treated intravenously with ViperaTAb or intramuscularly with European viper venom antiserum that was feasible only due to the unique situation of an antivenom shortage. The highest venom concentration, survival, length of hospital stay and adverse reactions did not differ between the groups. Patients treated with intravenous Fab fragments were sicker, with significantly more rhabdomyolysis and neurotoxicity. The kinetics of Fab fragments after one or more intravenous applications matched better with the venom concentration in the early phase of envenomation compared to F(ab’)2 fragments that were given intramuscularly only on admission. F(ab’)2 fragments given intramuscularly had 25-fold longer apparent total body clearance and 14-fold longer elimination half-time compared to Fab fragments given intravenously (2 weeks vs. 24 h, respectively). In V.ammodytes-envenomed patients, the intramuscular use of specific F(ab’)2 fragments resulted in a slow rise of antivenom serum concentration that demanded their early administration but without the need for additional doses for complete resolution of all clinical signs of envenomation. Intravenous use of paraspecific Fab fragments resulted in the immediate rise of antivenom serum concentration that enabled their use according to the clinical progress, but multiple doses might be needed for efficient therapy of thrombocytopenia due to venom recurrence, while the progression of rhabdomyolysis and neurotoxic effects of the venom could not be prevented.

scholarly journals The Ugly Side of Colchicine

2021 ◽  
Vol 9 ◽  
pp. 232470962110297
Author(s):  
Justin Cozza ◽  
Tuong Vi Cassandra Do ◽  
Shyam Ganti ◽  
Jayaramakrishna Depa
Keyword(s):  
Liver Failure ◽  
Acute Liver Failure ◽  
Activated Charcoal ◽  
The United States ◽  
Multiple Organ ◽  
Poison Control ◽  
Fab Fragments ◽  
Early Use ◽  
Potential Use ◽  
Electrolyte Abnormalities

We report a rare case of a 32-year-old male who ingested 32.4 to 54 mg of colchicine and presented after 44 hours. He developed progressive multiple organ failure with shock, acute kidney failure, troponemia, pancytopenia, absolute neutropenia, disseminated intravascular coagulation, acute liver failure, rhabdomyolysis, and lactic acidosis. He also developed electrolyte abnormalities and refractory hypoglycemia. Initial treatment consisted of activated charcoal, fluids, and broad-spectrum antibiotics with supportive treatment of mechanical ventilation, hemodialysis, vasopressors, N-acetylcysteine, colony-stimulating factors, and blood products. Literature shows potential benefit of colchicine-specific Fab fragments for acute toxicity with limited studies and is not currently available in the United States. Further research for N-acetylcysteine protocol for acute liver failure in colchicine toxicity and potential use of colchicine-specific Fab fragments is needed. Our case demonstrates the importance of early use of activated charcoal for ingestion overdose with the incorporation of poison control into multidisciplinary team for coordinated patient care.

scholarly journals Preliminary crystallographic study of the Fab fragments of two monoclonal anti-2-phenyloxazolone antibodies.

1985 ◽  
Vol 260 (18) ◽  
pp. 10268-10270
Author(s):  
R A Mariuzza ◽  
G Boulot ◽  
V Guillon ◽  
R J Poljak ◽  
C Berek ◽  
...  
Keyword(s):  
Fab Fragments ◽  
Crystallographic Study

scholarly journals Antibody in Middle Ear Fluid of Children Originates Predominantly from Sera and Nasopharyngeal Secretions

2012 ◽  
Vol 19 (10) ◽  
pp. 1593-1596 ◽  
Author(s):  
Ravinder Kaur ◽  
Thomas Kim ◽  
Janet R. Casey ◽  
Michael E. Pichichero
Keyword(s):  
Middle Ear ◽  
Acute Otitis Media ◽  
Secretory Iga ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Two Dimensional Gel Electrophoresis ◽  
Middle Ear Fluid ◽  
Iga Antibodies ◽  
Iga Antibody ◽  
Entire Array

ABSTRACTThe human middle ear is devoid of any immunocompetent cells in normal mucosa. We sought to determine the source of antibody present in the middle ear of children. Total IgG, IgA, and secretory IgA antibodies were determined by enzyme-linked immunosorbent assay from the nasopharyngeal, middle ear, and serum samples of children with acute otitis media. The two-dimensional gel electrophoresis pattern of the entire array of IgA antibodies in the nasal wash (NW) and middle ear fluid (MEF) was compared from the MEF and NW samples using isoelectric focusing and Western blotting. The total IgG and IgA antibodies in the MEF and NW samples of 137 children were compared. The ratio of IgG to IgA in the MEF was significantly different (P< 0.008) compared to NW because IgA levels were higher and IgG levels lower in NW. The IgG/IgA ratio of MEF resembled serum consistent with transudation to the MEF. Small amounts of secretory IgA were detected in MEF but the electrophoresis patterns of the entire array of IgA antibodies in the MEF and NW were virtually identical in each child evaluated; thus, IgA in MEF derived predominantly from serum and the nasopharynx by reflux via the Eustachian tube. The IgG/IgA antibody levels in the MEF and the same composition of IgA antibody in the MEF and NW identifies the predominant source of antibody in the MEF as a transudate of serum combined with nasal secretions refluxed from the nasopharynx in children.

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