With heart failure (HF) and renal malfunction becoming global public health issues, there is an urgent need to monitor diseases at home or in the community. Point-of-care testing (POC) would shorten the patients waiting time compared with laboratory molecular analysis. This work evaluates two types of gold nanomaterials, and two assay protocols, to develop a lateral flow (LF) system for Cystatin C (CysC) quantification. Of the protocols, the 'delayed-release' shows the alleviation of the hook effects with 1% BSA running buffer (RB), albeit at increased complexity with three steps of washing. The standard method with sample dilution (1: 150 sample dilution for GNPs, and 1:10 sample dilution for GNRs) can ensure the clinical range detection of CysC as 1 mg/L with partial LF assays. GNPs have stronger optical signal intensity compared with GNRs and developed full LF assays with GNPs require 1:1.5 sample dilution in recombinant Cys C detection. The ideal sample dilution ratio is different for partial and full LF assays. Clinical Relevance- This work is the basis of future work that will use LF devices for human serum/plasma monitoring to assess kidney function related to heart failure during medication. The specificity, sensitivity, and limit of detection will be validated via a clinical trial before potential clinical use.