Antibodies to liposomal phosphatidylserine and phosphatidic acid

1990 ◽  
Vol 68 (1) ◽  
pp. 96-101 ◽  
Author(s):  
Benoy Banerji ◽  
Carl R. Alving
Keyword(s):  
Phosphatidic Acid ◽  
Lipid A ◽  
Specific Activity ◽  
Cross Reactivity ◽  
Membrane Composition ◽  
Liposomal Membrane ◽  
Beef Brain ◽  
Polyclonal Antisera ◽  
High Degree

Polyclonal antisera to phosphatidylserine or phosphatidic acid were induced in rabbits by injecting liposomes containing phosphatidylserine or phosphatidic acid and lipid A. Adsorption of antisera with liposomes containing different phospholipids revealed that some degree of reactivity with one or more phospholipids other than the immunizing phospholipid was often observed. However, cross-reactivity with other phospholipids was not a universal phenomenon, and one antiserum to phosphatidylserine failed to cross-react (i.e., was not adsorbed) with liposomes containing other phospholipids. All of the antisera were inhibited by soluble phosphorylated haptens (e.g., phosphocholine but not choline), but one of the antisera to phosphatidylserine was inhibited both by phosphoserine and by serine alone. Liposomal membrane composition influenced the activity of antiserum to phosphatidylserine. Regardless of whether unsaturated (beef brain) or saturated (dimyristoyl) phosphatidylserine was used in the immunizing liposomes, the antisera reacted more vigorously with liposomes containing unsaturated than saturated phosphatidylserine. We conclude that liposomes containing lipid A can serve as vehicles for stimulating polyclonal antisera to phosphatidylserine and phosphatidic acid. Although cross-reactivity with certain other phospholipids can be observed, sera from selected animals apparently can exhibit a high degree of specific activity to the immunizing phospholipid antigen.Key words: liposomes, antibodies, phospholipids, phosphatidylserine, phosphatidic acid.

Antibodies to liposomal phosphatidylcholine and phosphatidylsulfocholine

1990 ◽  
Vol 68 (1) ◽  
pp. 54-64 ◽  
Author(s):  
Nabila M. Wassef ◽  
Glenn M. Swartz Jr. ◽  
Carl R. Alving ◽  
Morris Kates
Keyword(s):  
Immunosorbent Assay ◽  
Lipid A ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Activity ◽  
Dimyristoyl Phosphatidylcholine ◽  
Polyclonal Antisera ◽  
Mixed Populations ◽  
High Degree

Antibodies against dimyristoyl phosphatidylsulfocholine or dimyristoyl phosphatidylcholine were raised in rabbits after injection of liposomes containing phosphatidylsulfocholine or phosphatidylcholine, cholesterol, and lipid A. The antibody activities were assayed by complement-dependent immune damage to liposomes and by a solid-phase, enzyme-linked immunosorbent assay using purified dimyristoyl phosphatidylcholine or dimyristoyl phosphatidylsulfocholine as antigen. Each antiserum raised against phosphatidylsulfocholine reacted with liposomes containing phosphatidylcholine, and each antiserum raised against phosphatidylcholine reacted with liposomes containing phosphatidylsulfocholine. However, adsorption of dimyristoyl phosphatidylsulfocholine antiserum with liposomes containing dimyristoyl phosphatidylcholine removed all activity against dimyristoyl phosphatidylcholine, but did not eliminate antibody activity against dimyristoyl phosphatidylsulfocholine. These results indicate that the antiserum against phosphatidylsulfocholine contained mixed populations of antibodies. Polyclonal antisera that have been appropriately adsorbed can therefore be obtained with a high degree of specificity for phosphatidylsulfocholine and such antisera can distinguish between phosphatidylsulfocholine and phosphatidylcholine.Key words: liposomes, antibodies, phosphatidylsulfocholine, phosphatidylcholine.

scholarly journals High Enzymatic Recovery and Purification of Xylooligosaccharides from Empty Fruit Bunch via Nanofiltration

Processes ◽  
2020 ◽  
Vol 8 (5) ◽  
pp. 619
Author(s):  
Hans Wijaya ◽  
Kengo Sasaki ◽  
Prihardi Kahar ◽  
Nanik Rahmani ◽  
Euis Hermiati ◽  
...  
Keyword(s):  
Membrane Separation ◽  
Specific Activity ◽  
High Yield ◽  
Dual Function ◽  
Empty Fruit Bunch ◽  
Alkali Pretreatment ◽  
Nanofiltration Membranes ◽  
Degree Of Separation ◽  
Main Component ◽  
High Degree

Xylooligosaccharides (XOS) are attracting an ever-increasing amount of interest for use as food prebiotics. In this study, we used efficient membrane separation technology to convert lignocellulosic materials into a renewable source of XOS. This study revealed a dual function of nanofiltration membranes by first achieving a high yield of xylobiose (a main component of XOS) from alkali-pretreated empty fruit bunch (EFB) hydrolysate, and then by achieving a high degree of separation for xylose as a monosaccharide product. Alkali pretreatment could increase the xylan content retention of raw EFB from 23.4% to 26.9%, which eventually contributed to higher yields of both xylobiose and xylose. Nanofiltration increased the total amount of XYN10Ks_480 endoxylanase produced from recombinant Streptomyces lividans 1326 without altering its specific activity. Concentrated XYN10Ks_480 endoxylanase was applied to the recovery of both xylobiose and xylose from alkali-pretreated EFB hydrolysate. Xylobiose and xylose yields reached 41.1% and 17.3%, respectively, and when unconcentrated XYN10Ks_480 endoxylanase was applied, those yields reached 35.1% and 8.3%, respectively. The last step in nanofiltration was to separate xylobiose over xylose, and 41.3 g.L−1 xylobiose (90.1% purity over xylose) was achieved. This nanofiltration method should shorten the processes used to obtain XOS as a high-value end product from lignocellulosic biomass.

scholarly journals Characterization of Multiple forms of Human Thrombin

1977 ◽  
Author(s):  
J.J. Gorman ◽  
P.A. Castaldi
Keyword(s):  
Specific Activity ◽  
Peptide Mapping ◽  
Multiple Forms ◽  
Bovine Thrombin ◽  
Molecular Weights ◽  
Human Thrombin ◽  
Affinity Resin ◽  
C 50 ◽  
High Degree

Human thrombin was obtained by activation of partially purified human prothrombin with venom of the Australian Taipan (oxyuranus scutellatus scutellatus).The crude thrombin was precipitated with ammonium sulphate and subsequently purified by chromatography on Sephadex G-75 CM-Sephadex C-50 and the affinity resin am inobenzamidine-CH-Sepharose. The final preparation had a specific activity of 1700 units per absorbance unit (A| cm 280n m Was herterogenous as shown by urea-acrylamide gel electrophoresis at acid pH and by isoelectric focusing. SDS-acrylamide electrophoresis revealed molecular weights of 39,000, 28,000, 25-23,000 and 15-12,000 for these proteins. The 39,000 dalton species predominated (greater than 90%) when the enzyme was inhibited with phenyImethanesuI phony I fluoride prior to dialysis against 0.02M sod i urn phosphate (pH 8.0) containing 0.1% SDS. Lack of such inhibition reduced the amount of the 39,000 dalton species to less than 60% with concomitant increase in the smaller species. Increase in the smaller species also occurred during incubation in 0.IM NaCI-0.I M Tris buffer (pH 8.0).Peptide mapping studies indicated that the smaller species were structurally related to the 39,000 dalton species. Amino acid compositions of tryptic peptides indicated a high degree of homology with bovine thrombin.It has been established that human thrombin can exist in at least two secondary structural forms of different molecular weights, probably due to autolytic degradation of the largest (39,000 dalton) protein species.

PHOSPHORYLATION OF DIGLYCERIDES BY RAT BRAIN

1962 ◽  
Vol 40 (2) ◽  
pp. 247-259 ◽  
Author(s):  
K. P. Strickland
Keyword(s):  
Phosphatidic Acid ◽  
Rat Brain ◽  
Inorganic Phosphate ◽  
Specific Activity ◽  
Alkaline Treatment ◽  
Water Soluble ◽  
Two Dimensional ◽  
Relative Specific Activity ◽  
Hydrolysis Products ◽  
Stimulation Of

The addition of D-α,β-dimyristin was observed to stimulate by three to six times the labelling of phospholipids from radioactive inorganic phosphate (Pi32) by glycolysing homogenates and respiring mitochondria of rat brain. The increase in labelling was confined to the glycerophosphate (GP) isolated by two-dimensional chromatography from the water-soluble hydrolysis products obtained on weak alkaline treatment of the labelled phospholipids. The GP formed under these conditions is presumed to be derived mainly from phosphatidic acid formed by the phosphorylation of the diglyceride. A similar effect was observed for D-α,β-dipalmitin, D-α,β-diolein, and natural diglycerides prepared from either egg lecithin or spinal cord lecithin, but not for D-α-β-distearin. L-α,β-Diolein was much less effective than the D-isomer, suggesting a stereospecificity on the part of the enzymic phosphorylation of diglyceride. Experiments on the effects of the omission of Mg++ and the addition of glycolytic inhibitors on the stimulation of the labelling from Pi32 caused by D-α,β-dimyristin and D-α,β-diolein in the anaerobic homogenate system suggested that the increased phosphorylation caused by added diglycerides was closely coupled to active glycolysis. A comparison of the relative specific activity of the lipid P, following incubation of Pi32 and ATP32 in the anaerobic homogenate system inhibited by fluoride with and without D-α,β-diolein added, showed that the phosphate of the newly formed phosphatidic acid was derived from ATP, suggesting the presence of a D-α,β-diglyceride kinase.

scholarly journals Characterization of the Duffy-Binding-Like Domain of Plasmodium falciparum Blood-Stage Antigen 332

2011 ◽  
Vol 2011 ◽  
pp. 1-14 ◽  
Author(s):  
Sandra Nilsson ◽  
Kirsten Moll ◽  
Davide Angeletti ◽  
Letusa Albrecht ◽  
Inari Kursula ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Immunofluorescence Microscopy ◽  
Homology Model ◽  
Cross Reactivity ◽  
Blood Stage ◽  
Conserved Domain ◽  
Erythrocyte Binding ◽  
High Degree ◽  
Degree Of Similarity

Studies on Pf332, a major Plasmodium falciparum blood-stage antigen, have largely been hampered by the cross-reactive nature of antibodies generated against the molecule due to its high content of repeats, which are present in other malaria antigens. We previously reported the identification of a conserved domain in Pf332 with a high degree of similarity to the Duffy-binding-like (DBL) domains of the erythrocyte-binding-like (EBL) family. We here describe that antibodies towards Pf332-DBL are induced after repeated exposure to P. falciparum and that they are acquired early in life in areas of intense malaria transmission. Furthermore, a homology model of Pf332-DBL was found to be similar to the structure of the EBL-DBLs. Despite their similarities, antibodies towards Pf332-DBL did not display any cross-reactivity with EBL-proteins as demonstrated by immunofluorescence microscopy, Western blotting, and peptide microarray. Thus the DBL domain is an attractive region to use in further studies on the giant Pf332 molecule.

scholarly journals Cephalosporins’ Cross-Reactivity and the High Degree of Required Knowledge. Case Report and Review of the Literature

Antibiotics ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 209
Author(s):  
Stefano D’Errico ◽  
Paola Frati ◽  
Martina Zanon ◽  
Eleonora Valentinuz ◽  
Federico Manetti ◽  
...  
Keyword(s):  
Present Report ◽  
Specific Ige ◽  
Chain Structure ◽  
Cross Reactivity ◽  
Side Chain ◽  
Mast Cell Tryptase ◽  
Immunohistochemical Examination ◽  
Femoral Blood ◽  
The Subject ◽  
High Degree

Antibiotic cross-reactivity represents a phenomenon of considerable interest as well as antibiotic resistance. Immediate reactions to cephalosporins are reported in the literature with a prevalence of only 1–3% of the population, while anaphylactic reactions are rarely described (approximately 0.0001–0.1%) as well as fatalities. Allergic reaction to cephalosporins may occur because of sensitization to unique cephalosporin haptens or to determinants shared with penicillins. Cross-reactivity between cephalosporins represents, in fact, a well-known threatening event involving cephalosporins with similar or identical R1- or R2-side chains. The present report describes the case of a 79-year-old man who suddenly died after intramuscular administration of ceftriaxone. Serum dosage of mast cell tryptase from a femoral blood sample at 3 and 24 h detected values of 87.7μg/L and 93.5μg/L, respectively (cut-off value 44.3 μg/L); the serum-specific IgE for penicillins, amoxicillin, cephaclor and also for the most common allergens were also determined. A complete post-mortem examination was performed, including gross, histological and immunohistochemical examination, with an anti-tryptase antibody. The cause of death was identified as anaphylactic shock: past administrations of cefepime sensitized the subject to cephalosporins and a fatal cross-reactivity of ceftriaxone with cefepime occurred due to the identical seven-position side chain structure in both molecules. The reported case offers food for thought regarding the study of cross-reactivity and the need to clarify the predictability and preventability of the phenomenon in fatal events.

scholarly journals Cross reactivity of neutralizing antibodies to the encephalitic California Serogroup orthobunyaviruses varies by virus and genetic relatedness

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alyssa B. Evans ◽  
Karin E. Peterson
Keyword(s):  
Gold Standard ◽  
Neutralizing Antibodies ◽  
Viral Encephalitis ◽  
Genetic Relatedness ◽  
Cross Reactivity ◽  
Etiologic Agent ◽  
Snowshoe Hare ◽  
Neuroinvasive Disease ◽  
California Serogroup ◽  
High Degree

AbstractThe California Serogroup (CSG) of Orthobunyaviruses comprises several viruses capable of causing neuroinvasive disease in humans, including La Crosse (LACV), Snowshoe Hare (SSHV), Tahyna (TAHV), Jamestown Canyon (JCV), and Inkoo (INKV) viruses. Diagnosis of specific CSG viruses is complicated by the high degree of antibody cross-reactivity between them, with laboratory standards requiring a fourfold higher titer of neutralizating antibody (NAb) activity to positively identify the etiologic virus. To help elucidate NAb relationships between neuroinvasive CSG viruses, we directly compared the cross-reactivity of NAb between LACV, SSHV, TAHV, JCV, and INKV. Mice were inoculated with individual viruses and the NAb activity of plasma samples was compared by plaque reduction neutralization tests against all five viruses. Overall, the results from these studies show that the CSG viruses induced high levels of NAb against the inoculum virus, and differing amounts of cross-reactive NAb against heterologous viruses. LACV, SSHV, and INKV elicited the highest amount of cross-reactive NAb. Interestingly, a fourfold difference in NAb titer between the inoculum virus and the other CSG viruses was not always observed. Thus, NAb titers, which are the gold-standard for diagnosing the etiologic agent for viral encephalitis, may not clearly differentiate between different CSG viruses.

scholarly journals Genetic and antigenic diversity among noroviruses

2006 ◽  
Vol 87 (4) ◽  
pp. 909-919 ◽  
Author(s):  
Grant S. Hansman ◽  
Katsuro Natori ◽  
Haruko Shirato-Horikoshi ◽  
Satoko Ogawa ◽  
Tomoichiro Oka ◽  
...  
Keyword(s):  
Amino Acid ◽  
Insect Cells ◽  
Phylogenetic Analyses ◽  
Cross Reactivity ◽  
Amino Acid Sequences ◽  
Amino Acid Residues ◽  
Cell Culture Systems ◽  
Antibody Elisa ◽  
Polyclonal Antisera ◽  
Antigenic Relationships

Human norovirus (NoV) strains cause a considerable number of outbreaks of gastroenteritis worldwide. Based on their capsid gene (VP1) sequence, human NoV strains can be grouped into two genogroups (GI and GII) and at least 14 GI and 17 GII genotypes (GI/1–14 and GII/1–17). Human NoV strains cannot be propagated in cell-culture systems, but expression of recombinant VP1 in insect cells results in the formation of virus-like particles (VLPs). In order to understand NoV antigenic relationships better, cross-reactivity among 26 different NoV VLPs was analysed. Phylogenetic analyses grouped these NoV strains into six GI and 12 GII genotypes. An antibody ELISA using polyclonal antisera raised against these VLPs was used to determine cross-reactivity. Antisera reacted strongly with homologous VLPs; however, a number of novel cross-reactivities among different genotypes was observed. For example, GI/11 antiserum showed a broad-range cross-reactivity, detecting two GI and 10 GII genotypes. Likewise, GII/1, GII/10 and GII/12 antisera showed a broad-range cross-reactivity, detecting several other distinct GII genotypes. Alignment of VP1 amino acid sequences suggested that these broad-range cross-reactivities were due to conserved amino acid residues located within the shell and/or P1-1 domains. However, unusual cross-reactivities among different GII/3 antisera were found, with the results indicating that both conserved amino acid residues and VP1 secondary structures influence antigenicity.

scholarly journals Plasmid-mediated trimethoprim-resistance in Staphylococcus aureus. Characterization of the first gram-positive plasmid dihydrofolate reductase (type S1)

1987 ◽  
Vol 243 (1) ◽  
pp. 309-312 ◽  
Author(s):  
H K Young ◽  
R A Skurray ◽  
S G B Amyes
Keyword(s):  
Staphylococcus Aureus ◽  
Dihydrofolate Reductase ◽  
Specific Activity ◽  
Gram Positive ◽  
Heat Stable ◽  
Dihydrofolate Reductases ◽  
High Degree ◽  
Moderately Resistant ◽  
Identification And Characterization

The trimethoprim-resistance gene located on plasmid pSK1, originally identified in a multi-resistant Staphylococcus aureus from Australia, encodes the production of a dihydrofolate reductase (type S1), which confers a high degree of resistance to its host and is quite unlike any plasmid-encoded dihydrofolate reductase hitherto described. It has a low Mr (19,700) and has a higher specific activity than the constitutive Gram-negative plasmid dihydrofolate reductases. The type S1 enzyme is heat-stable and has a relatively low affinity for the substrate, dihydrofolate (Km 10.8 microM). It is moderately resistant to trimethoprim, and is competitively inhibited by this drug with an inhibitor-binding constant of 11.6 microM. This is the first identification and characterization of a plasmid-encoded trimethoprim-resistant dihydrofolate reductase derived from a Gram-positive species.

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