Solid-phase vitamin B12 assays using polyacrylamide-bound intrinsic factor and polyacrylamide-bound R-binder

Area of Science:

• Biochemistry
• Clinical Chemistry
• Hematology

Background:

• Vitamin B12 (cobalamin) deficiency is a significant cause of megaloblastic anemia.
• Accurate measurement of both total and microbiologically-available vitamin B12 is crucial for diagnosis.
• Existing assays may not always differentiate between various forms of vitamin B12 or its bioavailability.

Purpose of the Study:

• To develop and describe a new, simple solid-phase assay for vitamin B12.
• To simultaneously measure microbiologically-available vitamin B12 and total vitamin B12.
• To evaluate the assay's ability to differentiate between healthy controls and patients with vitamin B12 deficiency.

Main Methods:

• Development of a solid-phase reagent using intrinsic factor coupled to polyacrylamide beads for bioavailable B12.
• Development of a solid-phase reagent using salivary R-binder coupled to polyacrylamide beads for total B12.
• Application of the assay to sera from control subjects and patients with megaloblastic anemia due to vitamin B12 deficiency.

Main Results:

• The described assay is simple to perform.
• The assay successfully measures microbiologically-available vitamin B12 using intrinsic factor.
• The assay successfully measures total vitamin B12 using R-binder.
• The assay demonstrated complete separation of sera between control and patient groups.

Conclusions:

• The new solid-phase vitamin B12 assay is effective for measuring both bioavailable and total vitamin B12.
• This assay provides a clear diagnostic distinction for vitamin B12 deficiency.
• The simplicity and effectiveness of this assay make it a valuable tool in clinical diagnostics.