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The supernatants were filtered through 110 μm Waterman filter membrane and were mixed with adequate volumes of Ni-NTA agarose resin (5 PRIME, Inc., Gaithersburg, MD) for 1 hour at 4 °C

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Amidated peptides have been observed to exhibit reduced sensitivity to proteolytic degradation and, in some cases, increased receptor binding affinity relative to their acid-form counterparts (2)