Purified Mouse scFv Anti-Human CD45
Alternative Name: PTPRD; CD45R; LCA; L-CA; Leukocyte Common Antigen
Concentration: 0.5 mg/ml (Lot to Lot Variation)
Isotype: Mouse lgG1
Reactivity: Human Leukocytes
Application: ELISA Capture, Flow Cytometry, Immunofluorescence
Immunogen: Human Peripheral Blood Leukocytes
Storage Buffer: Aqueous buffered solution containing BSA and
≤ 0.09% sodium azide
Regulatory Status: Research Use Only
Sizes: 100 µg
Catalog No.: 01-SBTCD45scFv-2019
$100.00 per 100 µg
The HI30 monoclonal antibody specifically binds to the 180, 190, 205, 220 kDa protein isoforms of CD45. CD45 is encoded by the PTPRC (Protein tyrosine phosphatase receptor type C) gene. CD45, also known as the leukocyte common antigen (LCA), is a member of the protein tyrosine phosphatase (PTP) family. It is present on all human leukocytes including lymphocytes, monocytes, granulocytes, eosinophils, and thymocytes. CD45 is absent from circulating erythrocytes, platelets, or mature erythroid cells of bone marrow and non-hemopoietic tissues.
SBT has developed an anti-cd45 scFv antibody fragment (Figure below). This scFv fragment representing the functional antigen binding fragment of immunoglobulins is produced in bacterial expression systems. Using biochemical procedures and/or genetic modification (recombinant DNA technology), these fragments can be coupled to compounds and proteins that allow for the detection of these fragments. Examples of such compounds include fluorophores (e.g. fluorescein) and enzymes (e.g. alkaline phosphatase) generating fluorescence. The resultant compounds now have bifunctional properties in that they can bind to their target via the scFv fragment and can be detected via their fluorescence marker. In addition to labeling the scFv with fluorophore markers, SBT has coupled the scFv antibody fragment to magnetic microparticles allowing for the capture of the scFv target complex in a magnetic field.
An scFv fragment retains the complete antigen-binding capability of the whole monoclonal antibody, thus facilitating a unique molecule with the property of being used as either a diagnostic or therapeutic reagent. In addition, the scFv with exquisite specificity and affinity towards a specific target can also be used as a delivery vehicle for radioimmunoimaging and radioimmunotherapy. In addition, the small antigen-binding scFv offer several advantages over a whole antibody molecule in that the smaller size of the scFv allow it to penetrate more rapidly and evenly to tumors and can penetrate the blood brain-barrier. scFv fragments have more rapid clearance from blood, and therefore be coupled with drugs and radionuclides in order to result in low exposure of the healthy tissue. Also, there appears to be no uptake of the scFv by the kidney allowing them to efficiently localize to the tumors, an important property in cancer therapy.
Comparison of IgG and scFv
SBT developed a leukocyte depletion protocol, validating that the SBT CD45scFv performed as good if not better than a commercially available full length monoclonalCD45 antibody using fluorescein magnetic microparticles for leukocyte depletion by flow cytometry with human blood.
Polyacrylamide Gel Demonstrating Single Band of Purity of scFv anti cd45
PREPARATION AND STORAGE
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The scFv antibody fragment was purified from disrupted Escherichia coli culture by column chromatography.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
3. This reagent has been pre-diluted for use at the recommended volume per test. Typically, we use 1 × 10^6 cells in a 100-µl experimental sample (a test).
STABLEBODY REFERENCES FOR scFv anti-CD45
In collaboration with Rush University Medical Center, StableBody’s FITC Anti-Human CD45scFv conjugate using FACS was investigated on fresh human white blood cells and found it to work equivalently to the Beckman Coulter reagent Anti-Human CD45-APC.
In collaboration with IVDiagnostics LLC, StableBody’s FITC Anti-Human CD45scFv conjugate using FACS was investigated on fresh human white blood cells and found it to work equivalently to the BD/Pharmagen FITC Mouse Anti-Human CD45 reagent.
In collaboration with Spherotech Inc, scFv coated 1 micron magnetic beads showed excellent dose response (based on spectraMax and Flowcytometry signals) with 1 nanogram of Dylight labeled CD45 antigen. “Further information on fluorophore conjugation upon request.”
In a study done with the University of Wisconsin Comprehensive Cancer Center Flow Lab, SBT-CD45scFv conjugated to Dylight 650 showed results were concentration dependent when compared to a full length anti-CD45 antibody conjugated to a similar fluorophore (647 nm wavelength).