Construction and Evaluation of an Antibody Phage Display Library Targeting Heparan Sulfate

Construction and Evaluation of an Antibody Phage Display Library Targeting Heparan Sulfate
Heparan sulfate (HS) is a linear polysaccharide with excessive structural range. Totally different HS epitopes have been detected and localized utilizing single chain variable fragment (scFv) antibodies from a ‘single pot’ phage show library containing a randomized complementarity figuring out area of the heavy chain (CDR3).
On this examine, we created a brand new library containing anti-HS scFvs that every one harbor a dp-38 heavy chain section the place the CDR3 area was engineered to include the XBBXBX heparin binding consensus website (X = any amino acid, B = R, Okay or H).
The library contained ~1.73 × 106 distinctive antibodies and was biopanned in opposition to HS from a number of sources. The chosen antibodies have been sequenced and chemically/immunohistologically characterised. Numerous 67 anti-HS scFv antibodies have been chosen, of which 31 contained a XBBXBX CDR3 sequence.
There was a transparent choice for glycine on the first and proline on the fourth place of the CDR3. The sequence GZZP(R/Okay)X (Z = R, Okay or H, however can also include N, S, or Q) was unusually overrepresented. Chosen antibodies reacted with HS/heparin, however not with different glycosaminoglycans.
Antibodies reacted differentially with respect to N-, 2-O, or 6-O-desulfated heparin preparations, and confirmed distinct topologies of HS epitopes in rat kidney sections. The library could also be instrumental within the choice of a big pool of HS epitope-specific antibodies, and – since all antibodies differ solely of their 6 amino acid CDR area – could also be a instrument for a rational design of antibodies recognizing particular HS sulfation patterns.

Heparan sulfate proteoglycans (HSPGs) and chondroitin sulfate proteoglycans (CSPGs) perform as endocytic receptors for an internalizing anti-nucleic acid antibody.

A subset of monoclonal anti-DNA autoantibodies enters quite a lot of residing cells. Right here, we aimed to determine the endocytic receptors acknowledged by an internalizing anti-nucleic acid autoantibody, the 3D8 single-chain variable fragment (scFv).
We discovered that cell floor binding and internalization of 3D8 scFv have been inhibited markedly in soluble heparan sulfate (HS)/chondroitin sulfate (CS)-deficient or -removed cells and within the presence of soluble HS and CS. 3D8 scFv colocalized intracellularly with both HS proteoglycans (HSPGs) or CSPGs in HeLa cells.
3D8 scFv was co-endocytosed and co-precipitated with consultant particular person HSPG and CSPG molecules: syndecan-2 (a transmembrane HSPG), glypican-3 (a glycosylphosphatidylinositol (GPI)-anchored HSPG); CD44 (a transmembrane CSPG); and brevican (a GPI-anchored CSPG).
Collected knowledge point out that 3D8 scFv binds to the negatively charged sugar chains of each HSPGs and CSPGs and is then internalized together with these molecules, no matter how these proteoglycans are related to the cell membrane.
That is the primary examine to indicate that anti-DNA antibodies enter cells through each HSPGs and CSPGs concurrently. The information might support understanding of endocytic receptors that bind anti-DNA autoantibodies. The examine additionally gives perception into potential cell membrane targets for macromolecular supply.

Anti-heparan sulfate antibody and useful lack of glomerular heparan sulfate proteoglycans in lupus nephritis.

Background The aim of this examine was to guage the options of heparan sulfate proteoglycans (HSPGs) as agrins of the glomerular basement membrane (GBM) and circulating anti-heparan sulfate (HS) antibodies in lupus nephritis, evaluating titers among the many following teams: lupus nephritis (LN), non-renal lupus, non-lupus nephritis, and wholesome controls.
Strategies The stage of nephritis was decided based mostly on the kidney biopsy. Alcian blue staining and immunohistochemical (IHC) staining for agrin have been carried out for histological analysis of GBM HSPGs in regular glomeruli, non-lupus membranous glomerulonephritis (MGN), and lupus MGN.
The outcomes have been used for measurement of the serum anti-HS antibody titers utilizing an enzyme-linked immunosorbent assay (ELISA) within the following teams: 38 wholesome controls, 38 non-lupus nephritis, 37 non-renal lupus, and 38 LN. Outcomes Glomerulus HSPGs have been stained bluish-green alongside the GBM with Alcian blue.
Nevertheless, IHC staining in opposition to agrin was virtually utterly destructive within the lupus MGN group in contrast with the traditional and non-lupus MGN teams, which confirmed brown staining of GBM. The next stage of anti-HS IgG was detected in LN in contrast with different teams, respectively. Greater titers have been related to the presence of SLE and nephritis.
The next diploma of proteinuria normalized to glomerular filtration fee (eGFR) was noticed in affiliation with increased anti-HS antibody titers in LN. Conclusion This examine demonstrated a useful lack of GBM HSPGs and better ranges of circulating anti-HS antibodies as a attribute characteristic of lupus nephritis, suggesting their involvement within the pathogenesis of lupus nephritis and proteinuria.

Isolation of Antibodies to Heparan Sulfate on Glypicans by Phage Show.

Heparan sulfate (HS) performs an essential position in improvement and illness. It interacts with many progress components, chemokines, and different ligands recognized to be essential for cell progress, motility, and differentiation.
Nevertheless, isolating an antibody to HS in mice, rabbits, or people is troublesome because of the poor immunogenicity of HS. Phage show is a serious antibody engineering know-how that permits the choice of antibodies for poorly immunogenic or extremely conserved antigens.
This protocol incorporates detailed procedures for HS antigen preparation and isolation of a phage displayed human single-chain Fv (HS20) that binds HS on glypican-3 (GPC3), and evaluation of the chosen phage antibody. It’s conceivable that the procedures described on this protocol could also be relevant to the isolation of antibodies for quite a lot of HS molecules.
Construction and Evaluation of an Antibody Phage Display Library Targeting Heparan Sulfate

The technology and characterisation of neutralising antibodies in opposition to the Theiler’s murine encephalomyelitis virus (TMEV) GDVII capsid reveals the potential binding website of the host cell co-receptor, heparan sulfate.

The early phases of picornavirus capsid meeting and the host components concerned are poorly understood. Because the localisation of viral proteins in contaminated cells can present info on their perform, antibodies in opposition to purified Theiler’s murine encephalomyelitis virus (TMEV) GDVII capsids have been generated by immunisation of rabbits.
The resultant anti-TMEV capsid antibodies recognised a C-terminal area of VP1 however not VP2 or VP3 by Western evaluation. Examination of the websites of TMEV capsid meeting by oblique immunofluorescence and confocal microscopy confirmed that at 5h submit an infection, capsid sign was diffusely cytoplasmic with robust perinuclear staining and moved into giant punctate constructions from 6 to 8h submit an infection.
A plaque discount neutralisation assay confirmed that the anti-TMEV capsid antibodies however not anti-VP1 antibodies might neutralise viral an infection in vitro. The VP1 C-terminal residues recognised by the anti-TMEV capsid antibodies have been mapped to a loop on the capsid floor close to to the putative receptor binding pocket.
In silico docking experiments confirmed that the recognized TMEV co-receptor, heparan sulfate, interacts with residues of VP1 within the putative receptor binding pocket, residues of VP3 within the adjoining pit and residues of the adjoining VP1 C-terminal loop which is recognised by the anti-TMEV capsid antibodies.

Heparan Sulfate Proteoglycan (HSPG) Antibody (FITC)

20-abx273784
  • EUR 543.60
  • EUR 292.80
  • EUR 1545.60
  • EUR 744.00
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  • 100 ug
  • 10 ug
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  • 200 ug
  • 50 ug

Heparan Sulfate Proteoglycan (HSPG) Antibody (Biotin)

20-abx272006
  • EUR 510.00
  • EUR 276.00
  • EUR 1429.20
  • EUR 693.60
  • EUR 393.60
  • 100 ug
  • 10 ug
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Heparan Sulfate Proteoglycan 2 (HSPG2) Antibody

20-abx176806
  • EUR 526.80
  • EUR 159.60
  • EUR 1479.60
  • EUR 710.40
  • EUR 393.60
  • 100 ug
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Heparan Sulfate Proteoglycan 2 (HSPG2) Antibody

20-abx176807
  • EUR 1479.60
  • EUR 710.40
  • 1 mg
  • 200 ug

Heparan Sulfate Proteoglycan 2 (HSPG2) Antibody

20-abx212992
  • EUR 493.20
  • EUR 360.00
  • 100 ul
  • 50 ul

Heparan Sulfate Proteoglycan 2 (HSPG2) Antibody

20-abx213095
  • EUR 493.20
  • EUR 360.00
  • 100 ul
  • 50 ul

Heparan Sulfate (HS) ELISA Kit

abx364991-96tests 96 tests
EUR 848.4

Heparan Sulfate (HS) ELISA Kit

20-abx585370
  • EUR 9567.60
  • EUR 5095.20
  • EUR 1177.20
  • 10 × 96 tests
  • 5 × 96 tests
  • 96 tests

Heparan Sulfate (HS) ELISA Kit

abx513537-96tests 96 tests
EUR 801.6

Heparan sulfate ELISA Kit (OKEH02552)

OKEH02552 96 Wells
EUR 1028.4
Description: Description of target: Heparan sulfate (HS) is a linear polysaccharide found in all animal tissues. It occurs as a proteoglycan (HSPG) in which two or three HS chains are attached in close proximity to cell surface or extracellular matrix proteins. It is in this form that HS binds to a variety of protein ligands and regulates a wide variety of biological activities, including developmental processes, angiogenesis, blood coagulation and tumour metastasis. HS has been shown to serve as cellular receptor for a number of viruses including the respiratory syncytial virus.;Species reactivity: All;Application: ;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 2.345 ng/mL

Chicken heparan sulfate ELISA Kit

ELA-E0623d 96 Tests
EUR 1113.6

Recombinant Heparan Sulfate Proteoglycan (HSPG)

4-RPA565Hu01
  • EUR 507.07
  • EUR 259.20
  • EUR 1571.52
  • EUR 603.84
  • EUR 1087.68
  • EUR 415.20
  • EUR 3748.80
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Human Heparan Sulfate Proteoglycan expressed in: E.coli

anti-Heparan Sulfate Proteoglycan 2

YF-PA23929 50 ul
EUR 400.8
Description: Mouse polyclonal to Heparan Sulfate Proteoglycan 2

Mouse Anti Heparin/Heparan Sulfate Monoclonal Antibody

DMABT-54795MH 0.1 mg
EUR 1263.6

Heparan Sulfate Proteoglycan 2 (HSPG2) Antibody (Biotin)

20-abx272294
  • EUR 543.60
  • EUR 292.80
  • EUR 1579.20
  • EUR 744.00
  • EUR 410.40
  • 100 ug
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Heparan Sulfate Proteoglycan (HSPG) Polyclonal Antibody (Human)

4-PAA565Hu01
  • EUR 278.40
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  • EUR 688.80
  • EUR 346.80
  • EUR 249.60
  • 100ul
  • 10ml
  • 1ml
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Description: A Rabbit polyclonal antibody against Human Heparan Sulfate Proteoglycan (HSPG)

Anti-Heparan Sulfate Proteoglycan 2/HSPG2 Antibody

PB9277 100ug/vial
EUR 400.8

Bovine Heparan sulfate, HS ELISA Kit

CELI-66081b 96 Tests
EUR 1113.6

Chicken Heparan sulfate, HS ELISA Kit

CELI-66081c 96 Tests
EUR 1113.6

Canine Heparan sulfate, HS ELISA Kit

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General Heparan sulfate, HS ELISA Kit

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Human Heparan sulfate, HS ELISA Kit

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Mouse Heparan sulfate, HS ELISA Kit

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Porcine Heparan sulfate, HS ELISA Kit

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Rat Heparan sulfate, HS ELISA Kit

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Rabbit Heparan sulfate, HS ELISA Kit

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EUR 1113.6

Chicken heparan sulfate,HS ELISA Kit

CN-00806C1 96T
EUR 525.6

Chicken heparan sulfate,HS ELISA Kit

CN-00806C2 48T
EUR 346.8

Human heparan sulfate,HS ELISA kit

E01H1352-192T 192 tests
EUR 1524
Description: A competitive ELISA for quantitative measurement of Human heparan sulfate,HS in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human heparan sulfate,HS ELISA kit

E01H1352-48 1 plate of 48 wells
EUR 624
Description: A competitive ELISA for quantitative measurement of Human heparan sulfate,HS in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human heparan sulfate,HS ELISA kit

E01H1352-96 1 plate of 96 wells
EUR 822
Description: A competitive ELISA for quantitative measurement of Human heparan sulfate,HS in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Human heparan sulfate, HS ELISA Kit

CSB-E09585h-24T 1 plate of 24 wells
EUR 198
Description: Quantitativecompetitive ELISA kit for measuring Human heparan sulfate, HS in samples from serum, plasma, urine, tissue homogenates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.

Human heparan sulfate, HS ELISA Kit

1-CSB-E09585h
  • EUR 1080.00
  • EUR 6571.20
  • EUR 3480.00
  • 1 plate of 96 wells
  • 10 plates of 96 wells each
  • 5 plates of 96 wells each
Description: Quantitativecompetitive ELISA kit for measuring Human heparan sulfate, HS in samples from serum, plasma, urine, tissue homogenates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.

General HS/ Heparan sulfate ELISA Kit

E0039Ge 1 Kit
EUR 860.4

Human heparan sulfate,HS ELISA Kit

CN-04522H1 96T
EUR 565.2

Human heparan sulfate,HS ELISA Kit

CN-04522H2 48T
EUR 386.4

Goat heparan sulfate,HS ELISA kit

E06H1352-192T 192 tests
EUR 1524
Description: A competitive ELISA for quantitative measurement of Goat heparan sulfate,HS in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat heparan sulfate,HS ELISA kit

E06H1352-48 1 plate of 48 wells
EUR 624
Description: A competitive ELISA for quantitative measurement of Goat heparan sulfate,HS in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.

Goat heparan sulfate,HS ELISA kit

E06H1352-96 1 plate of 96 wells
EUR 822
Description: A competitive ELISA for quantitative measurement of Goat heparan sulfate,HS in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
These findings recommend that the anti-TMEV capsid antibodies neutralise virus an infection by stopping heparan sulfate from binding to the capsid. The antibodies produced on this examine are an essential instrument for additional investigating virus-host cell interactions important to picornavirus meeting.

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