This study examined the clothing wearing comfort of elderly women by electroencephalogram (EEG) and electrocardiogram (ECG) analyses. This study utilized 7 elderly individuals aged 65 or more. Two kinds of clothing ensemble (control and prototype) were used as experimental clothing. The control consisted of a general clothing ensemble and the prototype consisted of clothing that added an extra gap. Subjects wore the control or prototype from 9:00 to 21:30 and EEG and ECG signals were measured in the last 30 minutes. The EEG analysis showed that relative band power of a and α/high β were higher when they wore the prototype rather than the control. The ECG analysis showed that absolute band power of HF was higher; however, absolute band power of LF and LF/HF was lower when they wore the prototype rather than the control. Subjects felt less stressful and more comfortable when they wore the prototype. The results demonstrate the necessity to develop clothing in consideration of the body changes in elderly women. It is significant that the assessment of wearing comfort was aided by the use of EEG and ECG analysis in the field of clothing and textiles.