APOTCAsia Pacific Occupational Therapy Congress
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Serum contains 2 proteins (apoTC and HC) that have the potential to cross-react with a holoTC-specific antibody.
In the RIA, an immobilized TC-specific mAb captures the total TC (apoTC plus holoTC) in the sample; the vitamin [B.sub.12] bound by holoTC is then released and measured with a conventional competitive assay protocol for vitamin [B.sub.12].
When analyzed by gel filtration, no apoTC was identified in the sample treated with the mAb-containing microspheres, as shown in Fig.
No interference was observed with the following substances added to serum: hemoglobin (10 g/L), bilirubin (0.42 g/L), lipid (7.6 g/L), human HC (1 nmol/ L), and human apoTC (2.1 nmol/L).
[5] Nonstandard abbreviations: Cbl, cobalamin; TC, transcobalamin (previously transcobalamin II); HC, haptocorrin (previously transcobalamin I and III); holoTC, holo-transcobalamin; mAb, monoclonal antibody; apoTC, apotranscobalamin; SPR, surface plasmon resonance; holoHC, holo-haptocorrin; and PBS, phosphate-buffered saline.
We produced [B.sub.12] beads and removed apoTC before ELISA quantification (10) of the protein moiety of holoTC.
The amount of [B.sub.12] beads needed for removal of apoTC was determined from experiments where increasing amounts of beads were added to 100 [micro]L of serum (Fig.
The results are in agreement with previous data from analysis of holo- and apoTC (6, 8, 9).