In Fig. 5.2, the elution peak is narrow and tall. What does this most strongly indicate?

Question

BlackTom AI · Accepted Answer

A narrow, tall elution peak indicates the product is being desorbed and collected in a very small volume relative to the volume that was loaded — i.e., the capture step both binds and concentrates the target.

Relevant concepts:
- Affinity capture (Protein A) selectively binds antibody from a large load volume and releases it during elution in a much smaller volume, producing a high, narrow absorbance peak (concentration effect).
- Peak width is proportional to elution volume; peak height (at fixed total mass) increases as elution volume decreases.
- Dispersion, column exhaustion, and elution triggers have distinct signatures on a chromatogram.

Step-by-step reasoning:
1. Observe the chromatogram: the load phase spans roughly 5–15 CV (≈10 CV loaded), while the elution peak is sharp around ~16–17 CV with a width of ≈0.5 CV.  
2. Approximate concentration factor: if ~10 CV of feed containing the product is captured and then eluted into 0.5 CV, the concentration increases by ~10/0.5 = 20-fold (illustrative calculation showing how a small elution volume produces a tall peak).  
3. This pattern (large loaded volume, small elution volume) most directly points to concentration by capture/elution.

Why the correct answer is correct:
- Protein A binds the antibody across the large load volume and releases it in a focused elution step; the narrow high peak is the chromatographic manifestation of that concentration.

Why other options are incorrect:
- “Sharp peak proves no dispersion/holdup” — a sharp peak suggests low elution volume but does not prove absence of system dispersion; dispersion would broaden peaks, but absence of broadening alone isn’t proof of zero holdup.
- “Column was necessarily exhausted during load” — exhaustion would produce product in the flow-through during load (breakthrough), which is not evidenced by the chromatogram’s load trace.
- “Conductivity is the main trigger for elution” — Protein A elution is typically pH-driven (acidic elution); conductivity changes may accompany buffer changes but are not the principal trigger shown.

Conclusion: The narrow, tall elution peak most strongly indicates that Protein A capture also concentrates product (small elution volume), matching the provided answer.

In Fig. 5.2, the elution peak is narrow and tall. What does this most strongly indicate?单项选择题

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