An accurate method to locate of the flexion-extension (F-E) axis and longitudinal rotation (LR) axis of the tibiofemoral joint is required to accurately characterize tibiofemoral kinematics. A method was recently developed to locate these axes using an instrumented spatial linkage (ISL) (2012, “On the Estimate of the Two Dominant Axes of the Knee Using an Instrumented Spatial Linkage,” J. Appl. Biomech., 28(2), pp. 200–209). However, a more comprehensive error analysis is needed to optimize the design and characterize the limitations of the device before using it experimentally. To better understand the errors in the use of an ISL in finding the F-E and LR axes, our objectives were to (1) develop a method to computationally determine the orientation and position errors in locating the F-E and LR axes due to transducer nonlinearity and hysteresis, ISL size and attachment position, and the pattern of applied tibiofemoral motion, (2) determine the optimal size and attachment position of an ISL to minimize these errors, (3) determine the best pattern of pattern of applied motion to minimize these errors, and (4) examine the sensitivity of the errors to range of flexion and internal-external (I-E) rotation. A mathematical model was created that consisted of a virtual “elbow-type” ISL that measured motion across a virtual tibiofemoral joint. Two orientation and two position errors were computed for each axis by simulating the axis-finding method for 200 iterations while adding transducer errors to the revolute joints of the virtual ISL. The ISL size and position that minimized these errors were determined from 1080 different combinations. The errors in locating the axes using the optimal ISL were calculated for each of three patterns of motion applied to the tibiofemoral joint, consisting of a sequential pattern of discrete tibiofemoral positions, a random pattern of discrete tibiofemoral positions, and a sequential pattern of continuous tibiofemoral positions. Finally, errors as a function of range of flexion and I-E rotation were determined using the optimal pattern of applied motion. An ISL that was attached to the anterior aspect of the knee with 300-mm link lengths had the lowest maximum error without colliding with the anatomy of the joint. A sequential pattern of discrete tibiofemoral positions limited the largest orientation or position error without displaying large bias error. Finally, the minimum range of applied motion that ensured all errors were below 1 deg or 1 mm was 30 deg flexion with ±15 deg I-E rotation. Thus a method for comprehensive analysis of error when using this axis-finding method has been established, and was used to determine the optimal ISL and range of applied motion; this method of analysis could be used to determine the errors for any ISL size and position, any applied motion, and potentially any anatomical joint.