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There are several reasons for lower spike and recovery return values with Protein A ELISA kit (ADI-900-057). One possible reason could be that the protein A in the sample is higher than 1 mg/ml and this would result in not being fully dissociated upon boiling.
According to Step 1 in Sample Handling, we recommend to dilute all samples to 1 mg/mL in Assay Buffer 13 and incubate samples with buffer for 5 minutes in a boiling water bath. At higher sample concentration, 5 minutes would not be adequate to fully dissociate the protein A in samples. In addition, boiling conditions, concentration and time of incubation requires optimization depending with specific samples. For example, samples containing human IgG, an antibody with high affinity for Protein A, require dilution to 50 µg/mL and an additional 15 minutes of boiling to achieve accurate Protein A concentration.